QC Assay Validation
Transcript of QC Assay Validation
Sarthak Biotech Pvt. Ltd. Total Page: 01-06
Assay Method Validation QC/VLD/01
Name of the Product: SARCIP-500 TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 18/11/2007 Review Date : 17/11/2009 Revision No. : 00
OBJECTIVE : The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Ciprofloxacin in Sarcip-500 Tablet will give results with acceptable Accuracy, Linearity, Repeatibility, Relative Standard Deviation & Ruggedness when applied to the Sarcip-500 Tablet.
SCOPE : This method applies to the procedure which is used for calculation and assay of Ciprofloxacin.
RESPONSIBILITY: 1. Quality Control Manager.
2. Asstt. Quality Control Chemist.
VALIDATION PARAMETERS:
1. Linearity
2. Repeatibility.
3. Relative Standard Deviation.
4. Ruggedness.
Sarthak Biotech Pvt. Ltd. Total Page: 02-06Assay Method Validation QC/VLD/01Name of the Product: SARCIP-500 TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 18/11/2007Review Date : 17/11/2009 Revision No. : 00
METHOD :
1. Weight accurately equivalent to about 100mg of Ciprofloxacin working Reference standard and its equivalent to test in a 100ml volumetric flask . Dissolve with 0.1N- Hydrochloric Acid and make up to same, mix and filter if required.
2. With above dilution prepare standard and sample solution of 100mcg/ml with 0.1N Hydrochloric acid separately, mix and make homogeneous solution.
3. Pipette 3ml,5ml,7ml,9ml and 11ml of standard and sample solution separately in a calibrated 50ml.volumetric flask.
4. To each flask add 1ml of 5%w\v solution of Anhydrous Ferric chloride in water. Make up with distilled water to produced 50ml, mix to make homogeneous solution.
5. Measure the absorbance at maximum at about 438nm,against water as blank and tabulate the observed data in the observation table.
6. Plot linearity curve from the different concentration corresponding area of Ciprofloxacin peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.
7. Plot repeatability curve from by taking Ciprofloxacin solution serial no. on X-axis and observed peak area on Y-axis.
8. Draw a line which should cover almost all the points. The resulted graph should be
almost parallel to the X-axis. . .
9. Calculated the result by comparison with peak area obtained in standard and sample solution respectively.
Sarthak Biotech Pvt. Ltd. Total Page: 03-06Assay Method Validation QC/VLD/01Name of the Product: SARCIP-500 TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 18/11/2007Review Date : 17/11/2009 Revision No. : 00
LINEARITY STUDY
The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range.
Linearity of assay method of Ciprofloxacin in Sarcip-500 Tables shall be studied by measuring the absorbance of the standard Ciprofloxacin solution of 20 – 100mcg/ml concentration & range 438nm. Linearity curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR LINEARITY
Sr.No. Concentration of Ciprofloxacin in mcg\ml Peak area
1. 20mcg\ml 405
2. 40mcg\ml 450
3. 60mcg\ml 535
4. 80mcg\ml 570
5. 100mcg\ml 605
Sarthak Biotech Pvt. Ltd. Total Page: 04-06
Assay Method Validation QC/VLD/01
Name of the Product: SARCIP-500 TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 18/11/2007 Review Date : 17/11/2009 Revision No. : 00
REPEATIBILITY STUDY
Repeatibility of assay method of Ciprofloxacin in Sarcip-500 Tables shall be studied by measuring the absorbance of the standard Ciprofloxacin solution of 100mcg/ml concentration & range 438nm. Repeatibility curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR REPEATIBILITY
Sr. No. Concentration in mcg\ml Peak area 1 100mcg\ml 604
2 . 100mcg\ml 608
3. 100mcg\ml 603
4. 100mcg\ml 605
5. 100mcg\ml 604
Sarthak Biotech Pvt. Ltd. Total Page: 05-06
Assay Method Validation QC/VLD/01Name of the Product: SARCIP-500 TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 18/11/2007Review Date : 17/11/2009 Revision No. : 00
RELATIVE STANDARD DEVIATION
Test concentration of Ciprofloxacin used = 100mcg\ml.
Number of test solution used = 5 604+608+603+605+604 Average area = ----------------------------------- = 604.8
5
(604-604.8)2 +(608-604.8)2 +(603-604.8)2 +(605-604.8)2 +(604-604.8)2
Dev. S = -------------------------------------------------------------------------------------- 1/2
5-1
0.64+10.24+3.24+0.04+0.64 = --------------------------------------1/2
4
14.8 = -------1/2
4
= 3.7 1/2
= 1.9235
1.9235 x 100 RSD = ------------------ = 0.318%
604.8
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd. Total Page: 06-06Assay Method Validation QC/VLD/01Name of the Product: SARCIP-500 TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 18/11/2007 Review Date : 17/11/2009 Revision No. : 00
RUGGEDNESS STUDY
The ruggedness of an analytical method is the degree of reproducibility of the test
results obtained by the analysis of the samples from same lot under a variety of normal test
conditions (i.e. different laboratories, analysts, days & Instruments).
The ruggedness of assay method of Ciprofloxacin in Sarcip-500 tablet shall be studied
by analyzing the samples from same lot by two different analysts from same laboratory or at
two different laboratories by two different analysts on two different days.
RUGGEDNESS STUDY DATA
Ruggedness data for assay method by two different analysts on different days and different instrument.
Sr. No.
Analyst- I (Rajnish Kohli)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Analyst- II (Satpal Saini)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Wt. of Ref.
Standardgm.
Wt. of Sample
gm.
Measured Absor-
bance at of Ref.
Std.
Measured Absor-
bance at 438nm of
sample
% of Cipro-
floxacin
Wt. of Ref.
Standardgm.
Wt. of Sample
gm
Measured Absor-
bance at of Ref.
Std.
Measured Absor-
bance at 438nm
of sample
% of Cipro-
floxacin
01 0.1168 0.1348 602 608 100.11 0.1162 0.1349 606 601 99.43
02 0.1156 0.1352 607 612 99.61 0.1165 0.1356 609 616 101.15
03 0.1158 0.1342 605 615 101.35 0.1164 0.1352 612 608 99.55
Average wt of tablet 670.2 mg Averagewt.of tablet 675.1 mg
Mean 100.35% Mean 100.04%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 100.2 %Standard Deviation : ±10 %
Sarthak Biotech Pvt. Ltd. Total Page: 01-06Assay Method Validation QC/VLD/02Name of the Product: SARFLOK-200 TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 18/11/2007 Review Date : 17/11/2009 Revision No. : 00
PURPOSE :-The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Ofloxacin in Sarflok-200 Tablet will give results with acceptable Accuracy, Linearity, Repeatibility, Relative Standard Deviation & Ruggedness when applied to the Sarflok-200 Tablet.
SCOPE:- This method applies to the procedure which is used for calculation and assay of Ofloxacin.
RESPONSIBILITY: 1. Quality Control Manager.
2. Asstt. Quality Control Chemist.
VALIDATION PARAMETERS :-
1. Linearity.
2. Repeatibility.
3. Relative Standard Deviation.
4. Ruggedness.
Sarthak Biotech Pvt. Ltd. Total Page: 02-06Assay Method Validation QC/VLD/02Name of the Product: SARFLOK-200 TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 18/11/2007 Review Date : 17/11/2009 Revision No. : 00
METHOD
1. Weigh accurately equivalent to about 50mg of Ofloxacin working Reference standard and its equivalent to test in a 100ml volumetric flask . Dissolve with 0.1N- Hydrochloric Acid and make up to same, mix and filter if required.
2. Pipette 3ml,4ml,5ml,6ml and 7ml of standard and sample solution separately in a calibrated 50ml.volumetric flask.
4. To each flask add 1ml of 5%w\v solution of Ferric Chloride. Make up the volume with distilled water to produce 50ml, mix to make homogeneous solution.
5. Measure the absorbance at maximum at about 437nm,against water as blank and calculate the observed data in the observation table.
6. Plot linearity curve from the different concentration corresponding area of Ofloxacin peak. Taking concentration on X-axis and corresponding peak on Y-axis .Plot graph should be connected at least three points.
7. Plot repeatability curve from by taking Ofloxacin solution serial no. on X-axis and observed peak area on Y-axis.
8. Draw a line which should cover almost all the points. The resulted graph should be almost parallel to the X-axis. . .
9. Calculated the result by comparison with peak area obtained in standard and sample solution respectively.
Sarthak Biotech Pvt. Ltd. Total Page: 03-06Assay Method Validation QC/VLD/02Name of the Product: SARFLOK-200 TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 18/11/2007 Review Date : 17/11/2009 Revision No. : 00
LINEARITY STUDY
The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range.
Linearity of assay method of Ofloxacin in Sarflox-200 Tables shall be studied by measuring the absorbance of the standard Ofloxacin solution of 30 – 70mcg/ml concentration & range 437nm. Linearity curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR LINEARITY
Sr.No. Concentration of Ofloxacin in mcg\ml Peak area
1. 30mcg\ml 368
2. 40mcg\ml 437
3. 50mcg\ml 512
4. 60mcg\ml 591
5. 70mcg\ml 664
Sarthak Biotech Pvt. Ltd. Total Page: 04-06Assay Method Validation QC/VLD/02
Name of the Product: SARFLOK-200 TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 18/11/2007 Review Date : 17/11/2009 Revision No. : 00
REPEATIBILITY STUDY
Repeatibility of assay method of Ofloxacin in Sarflox-200 Tables shall be studied by measuring the absorbance of the standard Ofloxacin solution of 50 mcg/ml concentration & range 437nm.Repeatibility curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR REPEATIBILITY
S.No. Concentration in mcg\ml Peak area 1 50mcg\ml 515
2 . 50mcg\ml 512
3. 50mcg\ml 515
4. 50mcg\ml 509
5. 50mcg\ml 519
Sarthak Biotech Pvt. Ltd. Total Page: 05-06Assay Method Validation QC/VLD/02Name of the Product: SARFLOK-200 TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 18/11/2007 Review Date : 17/11/2009 Revision No. : 00
RELATIVE STANDARD DEVIATION
Test concentration of Ofloxacin used = 50mcg\ml. Number of test solution used = 5 515 + 512 + 515 + 509 + 519 Average area = ------------------------------------------ = 513.4
5
(515-513.4)2 +(512-513.4)2 +(515-513.4)2 +(509-513.4)2 +(519-513.4)2 1/2
Dev. S = --------------------------------------------------------------------------------------- 5-1
2.56+1.96+1.96+19.36+31.36 = ---------------------------------------1/2
4
57.2 = ------- 1/2
4
= 14.3 1/2 = 3.781
3.781 x 100 RSD = ------------------ = 0.736%
513.4
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd. Total Page: 06-06Assay Method Validation QC/VLD/02Name of the Product: SARFLOK-200 TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 18/11/2007 Review Date : 17/11/2009 Revision No. : 00
RUGGEDNESS STUDY
The ruggedness of an analytical method is the degree of reproducibility of the test
results obtained by the analysis of the samples from same lot under a variety of normal test
conditions (i.e. different laboratories, analysts, days & Instruments).
The ruggedness of assay method of Ofloxacin in Sarflok-200 tablet shall be studied by
analyzing the samples from same lot by two different analysts from same laboratory or at two
different laboratories by two different analysts on two different days.
RUGGEDNESS STUDY DATA
Ruggedness data for assay method by two different analysts on different days and different instrument.
Sr. No.
Analyst- I (Satpal Saini)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Analyst- II (Rajnish Kohli)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Wt. of Ref.
Standardgm.
Wt. of Sample
gm.
Measured Absorbanc
e at of Ref.
Std.
Measured Absorbance
at 437nm of sample
% of Ofloxacin
Wt. of Ref.
Standardgm.
Wt. of Sample
gm
Measured Absorbanc
e at of Ref.
Std.
Measured Absor-
bance at 437nm
of sample
% of Ofloxacin
01 0.0498 0.1021 509 512 100.67 0.0499 0.1006 512 505 98.97
02 0.0501 0.1019 511 515 101.7 0.0512 0.1018 504 510 102.95
03 0.0512 0.1029 519 512 100.7 0.0489 0.1012 508 514 98.90
Average wt of tablet 410.4 mg Averagewt.of tablet 404.6 mg
Mean 101.02% Mean 100.27%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 100.65 %Standard Deviation : ±10 %
Sarthak Biotech Pvt. Ltd. Total Page: 01-11Assay Method Validation QC/VLD/03Name of the Product: SARNIM-P TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 19/12/2007 Review Date : 18/12/2009 Revision No. : 00
PURPOSE : The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Nimesulide & Paracetamol in Sarnim-P Tablet will give results with acceptable Accuracy, Linearity, Repeatibility, Relative Standard Deviation & Ruggedness when applied to the Sarnim-P Tablet.
SCOPE : This method applies to the procedure which is used for calculation and assay of Paracetamol & Nimesulide.
RESPONSIBILITY : 1. Quality Control Manager.
2. Asstt. Quality Control Chemist.
VALIDATION PARAMETERS:
1. Linearity.
2. Repeatibility.
3. Relative Standard Deviation.
4. Ruggedness.
Sarthak Biotech Pvt. Ltd. Total Page: 02-11Assay Method Validation QC/VLD/03Name of the Product: SARNIM-P TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 19/12/2007 Review Date : 18/12/2009 Revision No. : 00
PROCEDURE : (For Paracetamol)
1. Weight accurately equivalent to about 60mg of Paracetamol working Reference standard and its equivalent to sample in a 100ml volumetric flask . Dissolve with 20ml of Hydrochloric Acid 1M and digest for 30 minutes, then make up the volume 100ml with distilled water.
2. Pipette 2ml,3ml,4ml,5ml and 6ml of standard and sample solution separately in a calibrated 50ml. measuring flask.
3. To each flask add 2ml of 10%w\v solution of Sodium Nitrate & allow to stand for 5 minutes, then add 2ml of 10% w/v solution of Ammonium Sulphamate. Again allow to stand for 5 to 6 minutes now further add 2 ml. of 10%w/v solution of Sodium Hydroxide and keep it for 5 minutes. Now adjust the volume to 50ml with distilled water.
4. Measure the absorbance at maximum at about 450nm, against water as blank and tabulate the observed data in the observation table.
5. Plot linearity curve from the different concentration corresponding area of Paracetamol peak. Taking concentration on X-axis and corresponding peak on Y-axis .Plot graph should be connected at least three points.
6. Plot repeatability curve from by taking Paracetamol solution serial no. on X-axis and
observed peak area on Y-axis.
7. Draw a line which should cover almost all the points. The resulted graph should be almost parallel to the X-axis. . .
8. Calculated the result by comparison with peak area obtained in standard and sample solution respectively.
Sarthak Biotech Pvt. Ltd. Total Page: 03-11Assay Method Validation QC/VLD/03Name of the Product: SARNIM-P TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 19/12/2007 Review Date : 18/12/2009 Revision No. : 00
LINEARITY STUDY
The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range.
Linearity of assay method of Paracetamol in Sarnim-P Tables shall be studied by measuring the absorbance of the standard Sarnim-P solution of 24 – 72mcg/ml concentration & range 450nm. Linearity curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR LINEARITY
Sr.No. Concentration of Paracetamol in mcg\ml Peak area
1. 24mcg\ml 525
2. 36mcg\ml 595
3. 48mcg\ml 645
4. 60mcg\ml 695
5. 72mcg\ml 742
Sarthak Biotech Pvt. Ltd. Total Page: 04-11Assay Method Validation QC/VLD/03Name of the Product: SARNIM-P TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 19/12/2007 Review Date : 18/12/2009 Revision No. : 00
REPEATIBILITY STUDY Repeatibility of assay method of Paracetamol in Sarnim-P Tables shall be studied by measuring the absorbance of the standard Sarnim-P solution of 24 mcg/ml concentration & range 450nm. Repeatibility curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR REPEATBILITY
Sr.No. Concentration in mcg\ml Peak area 1 24mcg\ml 525
2 . 24mcg\ml 528
3. 24mcg\ml 527
4. 24mcg\ml 526
5. 24mcg\ml 525
Sarthak Biotech Pvt. Ltd. Total Page: 05-11Assay Method Validation QC/VLD/03Name of the Product: SARNIM-P TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 19/12/2007 Review Date : 18/12/2009 Revision No. : 00
RELATIVE STANDARD DEVIATION
Test concentration of Paracetamol used = 24mcg\ml. Number of test solution used = 5 525 + 528 + 527 + 526 + 525 Average area = ------------------------------------------ = 526.2
5
(525-526.2)2 +(528-526.2)2 +(527-526.2)2 +(526-526.2)2 +(525-526-2)2 1/2
Dev. S = --------------------------------------------------------------------------------------- 5-1
1.44+3.24 +0.64+0.04+1.44 = ---------------------------------------1/2
4
6.8 = -------1/2
4
= 1.7 1/2 = 1.303
1.303 x 100 RSD = ------------------ = 0.2477%
526.2
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd. Total Page: 06-11Assay Method Validation QC/VLD/03Name of the Product: SARNIM-P TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 19/12/2007 Review Date : 18/12/2009 Revision No. : 00
RUGGEDNESS STUDY
The ruggedness of an analytical method is the degree of reproducibility of the test
results obtained by the analysis of the samples from same lot under a variety of normal test
conditions (i.e. different laboratories, analysts, days & Instruments).
The ruggedness of assay method of Paracetamol in Sarnim-P tablet shall be studied by
analyzing the samples from same lot by two different analysts from same laboratory or at two
different laboratories by two different analysts on two different days.
RUGGEDNESS STUDY DATA
Ruggedness data for assay method by two different analysts on different days and different instrument.
Sr. No.
Analyst- I (Satpal Saini)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Analyst- II (Rajnish Kohli)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Wt. of Ref.
Standardgm.
Wt. of Sample
gm.
Measured Absor-
bance at of Ref.
Std.
Measured Absor-
bance at 450nm of
sample
% of Paracetamol-mol
Wt. of Ref.
Standardgm.
Wt. of Sample
gm
Measured Absor-
bance at of Ref.
Std.
Measured Absor-
bance at 450nm
of sample
% of Paraceta-
mol
01 0.0598 0.0784 521 528 100.83 0.0602 0.0782 512 518 101.99
02 0.0601 0.0792 532 528 98.23 0.0592 0.0779 519 515 98.75
03 0.0592 0.0784 518 525 99.19 0.0612 0.0791 522 525 101.9
Average wt of tablet 652.2 mg Averagewt.of tablet 654.8 mg
Mean 99.41% Mean 100.88%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 100.15 %Standard Deviation : ±10 %
Sarthak Biotech Pvt. Ltd. Total Page: 07-11Assay Method Validation QC/VLD/03Name of the Product: SARNIM-P TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 19/12/2007 Review Date : 18/12/2009 Revision No. : 00
PROCEDURE : (For Nimesulide)
1. Weight accurately equivalent to about 50mg of Nimesulide working Reference standard and its equivalent to sample in a 100ml volumetric flask . Dissolve with 30 to 40ml of 0.1M solution of Sodium Hydroxide. Digest both the solution for 10 minutes then make up the volume to 100ml with 0.1M solution of Sodium Hydroxide.
2. Pipette 2ml,3ml,4ml,5ml and 6ml of standard and sample solution separately in a calibrated 50ml.measuring flask.
3. To each flask add 20ml 0.1M Sodium Hydroxide solution shake well and further adjust the volume up to 50ml with 0.1M solution of Sodium Hydroxide.
4. Measure the absorbance of both the solution at 405nm, against 0.1M Sodium Hydroxide solution as blank and tabulate the observed data in the observation table.
5. Plot linearity curve from the different concentration corresponding area of Nimesulide peak. Taking concentration on X-axis and corresponding peak on Y-axis .Plot graph should be connected at least three points.
6. Plot repeatability curve from by taking Nimesulide solution serial no. on X-axis and
observed peak area on Y-axis.
7. Draw a line which should cover almost all the points. The resulted graph should be almost parallel to the X-axis. . .
8. Calculated the result by comparison with peak area obtained in standard and sample solution respectively.
Sarthak Biotech Pvt. Ltd. Total Page: 08-11Assay Method Validation QC/VLD/03Name of the Product: SARNIM-P TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 19/12/2007 Review Date : 18/12/2009 Revision No. : 00
LINEARITY STUDY
The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range.
Linearity of assay method of Nimesulide in Sarnim-P Tablets shall be studied by measuring the absorbance of the standard Nimesulide solution of 20 – 60mcg/ml concentration & range 405nm. Linearity curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR LINEARITY
Sr.No. Concentration of Nimesulide in mcg\ml Peak area
1. 20mcg\ml 552
2. 30mcg\ml 595
3. 40mcg\ml 635
4. 50mcg\ml 702
5. 60mcg\ml 757
Sarthak Biotech Pvt. Ltd. Total Page: 09-11Assay Method Validation QC/VLD/03Name of the Product: SARNIM-P TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 19/12/2007 Review Date : 18/12/2009 Revision No. : 00
REPEATBILITY STUDY Repeatibility assay method of Nimesulide in Sarnim-P Tablets shall be studied by measuring the absorbance of the standard Nimesulide solution of 20 mcg/ml concentration & range 405nm. Repeatibility curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR REPEATIBILITY
Sr.No. Concentration in mcg\ml Peak area 1 20mcg\ml 559
2 . 20mcg\ml 561
3. 20mcg\ml 561
4. 20mcg\ml 560
5. 20mcg\ml 563
Sarthak Biotech Pvt. Ltd. Total Page: 10-11Assay Method Validation QC/VLD/03Name of the Product: SARNIM-P TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 19/12/2007 Review Date : 18/12/2009 Revision No. : 00
RELATIVE STANDARD DEVIATION
Test concentration of Nimesulide used = 20mcg\ml. Number of test solution used = 5 559 + 561 + 561 + 560 + 563 Average area = ------------------------------------------ = 560.8
5
(559-560.8)2 +(561-560.8)2 +(561-560.8)2 +(560-560.8)2 +(563-560.8)2 1/2
Dev. S = --------------------------------------------------------------------------------------- 5-1
3.24+0.04 +0.04+0.64+4.84 = ---------------------------------------1/2
4 8.8 = -------1/2
4
= 2.2 1/2 = 1.4142 1.4142 x 100 RSD = ------------------ = 0.2521%
560.8
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd. Total Page: 11-11Assay Method Validation QC/VLD/03Name of the Product: SARNIM-P TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 19/12/2007 Review Date : 18/12/2009 Revision No. : 00
RUGGEDNESS STUDY
The ruggedness of an analytical method is the degree of reproducibility of the test
results obtained by the analysis of the samples from same lot under a variety of normal test
conditions (i.e. different laboratories, analysts, days & Instruments).
The ruggedness of assay method of Nimesulide in Sarnim-P tablet shall be studied by
analyzing the samples from same lot by two different analysts from same laboratory or at two
different laboratories by two different analysts on two different days.
RUGGEDNESS STUDY DATA
Ruggedness data for assay method by two different analysts on different days and different instrument.
Sr. No.
Analyst- I (Rajnish Kohli)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Analyst- II (Satpal Saini)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Wt. of Ref.
Standardgm.
Wt. of Sample
gm.
Measured Absor-
bance at of Ref.
Std.
Measured Absor-
bance at 405nm of
sample
% of Nimesuli
de
Wt. of Ref.
Standardgm.
Wt. of Sample
gm
Measured Absor-
bance at of Ref.
Std.
Measured Absor-
bance at 405nm
of sample
% of Nimesuli
de
01 0.0499 0.3258 560 568 101.31 0.0498 0.3261 565 560 99.11
02 0.0489 0.3251 542 545 98.64 0.0496 0.3269 559 562 99.88
03 0.0501 0.3251 560 555 99.61 0.0502 0.3251 553 556 101.65
Average wt of tablet 652.2 mg Averagewt.of tablet 654.8 mg
Mean 99.85% Mean 100.21%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 100.03 %Standard Deviation : ±10 %
Sarthak Biotech Pvt. Ltd. Total Page: 01-06Assay Method Validation QC/VLD/04Name of the Product: SARFIX-100 DT TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 28/11/2007 Review Date : 27/11/2009 Revision No. : 00
PURPOSE :- The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Cefixime Trihydrate in Sarfix-100 DT Tablet will give results with acceptable Accuracy, Linearity, Repeatability, Relative Standard Deviation & Ruggedness when applied to the Sarfix-100 DT Tablet.
SCOPE:- This method applies to the procedure which is used for calculation and assay of Cefixime Trihydrate.
RESPONSIBILITY: 1. Quality Control Manager.
2. Asstt. Quality Control Chemist.
VALIDATION PARAMETERS :-
1. Linearity.
2. Repeatibility.
3. Relative Standard Deviation.
4. Ruggedness.
Sarthak Biotech Pvt. Ltd. Total Page: 02-06Assay Method Validation QC/VLD/04Name of the Product: SARFIX-100 DT TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 28/11/2007 Review Date : 27/11/2009 Revision No. : 00
METHOD
1. Weigh accurately equivalent to about 100mg of Cefixime working Reference standard and its equivalent to test in a 100ml volumetric flask . Dissolve with Methanol and make up to volume up to 100ml.
2. Pipette 1ml,2ml,3ml,4ml and 5ml of standard and sample solution separately in a calibrated 100ml.volumetric flask.
3. To each flask add Methanol and make up the volume up to 100ml.
4. Measure the absorbance at maximum at about 268nm,against Methanol as blank and calculate the observed data in the observation table.
5. Plot linearity curve from the different concentration corresponding area of Cefixime peak. Taking concentration on X-axis and corresponding peak on Y-axis .Plot graph should be connected at least three points.
6. Plot repeatability curve from by taking Cefixime solution serial no. on X-axis and observed peak area on Y-axis.
7. Draw a line which should cover almost all the points. The resulted graph should be
almost parallel to the X-axis. . .
8. Calculated the result by comparison with peak area obtained in standard and sample solution respectively.
Sarthak Biotech Pvt. Ltd. Total Page: 03-06Assay Method Validation QC/VLD/04Name of the Product: SARFIX-100 DT TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 28/11/2007 Review Date : 27/11/2009 Revision No. : 00
LINEARITY STUDY
The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range.
Linearity of assay method of Cefixime Trihydrate in Sarfix-100 DT Tablets shall be studied by measuring the absorbance of the standard Cefixime Trihydrate solution of 20 – 50mcg/ml concentration & range 268nm. Linearity curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR LINEARITY
Sr.No. Concentration of Cefixime in mcg\ml Peak area
1. 10mcg\ml 495
2. 20mcg\ml 515
3. 30mcg\ml 540
4. 40mcg\ml 570
5. 50mcg\ml 595
Sarthak Biotech Pvt. Ltd. Total Page: 04-06Assay Method Validation QC/VLD/04Name of the Product: SARFIX-100 DT TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 28/11/2007 Review Date : 27/11/2009 Revision No. : 00
REPEATIBILITY STUDY
Repeatibility of assay method of Cefixime Trihydrate in Sarfix-100 DT Tablets shall be studied by measuring the absorbance of the standard Cefixime Trihydrate solution of 10mcg/ml concentration & range 268nm. Repeatibility curve shall be plotted for absorbance against concentration
OBSERVATION TABLE FOR REPEATIBILITY
S.No. Concentration in mcg\ml Peak area 1 10mcg\ml 495
2 . 10mcg\ml 492
3. 10mcg\ml 497
4. 10mcg\ml 495
5. 10mcg\ml 496
Sarthak Biotech Pvt. Ltd. Total Page: 05-06Assay Method Validation QC/VLD/04Name of the Product: SARFIX-100 DT TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 28/11/2007 Review Date : 27/11/2009 Revision No. : 00
RELATIVE STANDARD DEVIATION Test concentration of Cefixime used = 10mcg\ml.
Number of test solution used = 5
495 + 492 + 497 + 495 + 496 Average area = ------------------------------------------ = 495
5
(495-495)2 +(495-492)2 +(495-497)2 +(495-495)2 +(495-496)2 1/2
Dev. S = -------------------------------------------------------------------------- 5-1
0+9+4+0+1 = ---------------------------------------1/2
4
14 = ------- 1/2
4
= 3.5 1/2
= 1.8708
1.8708 x 100 RSD = ------------------ = 0.377%
495
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd. Total Page: 06-06Assay Method Validation QC/VLD/04Name of the Product: SARFIX-100 DT TABLETPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 28/11/2007 Review Date : 27/11/2009 Revision No. : 00
RUGGEDNESS STUDY
The ruggedness of an analytical method is the degree of reproducibility of the test results obtained by the analysis of the samples from same lot under a variety of normal test
conditions (i.e. different laboratories, analysts, days & Instruments).The ruggedness of assay method of Cefixime Trihydrate in Sarfix-100 DT Tablet shall
be studied by analyzing the samples from same lot by two different analysts from same
laboratory or at two different laboratories by two different analysts on two different days.
RUGGEDNESS STUDY DATA
Ruggedness data for assay method by two different analysts on different days and different instrument.
Sr. No.
Analyst- I (Satpal Saini)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Analyst- II (Rajnish Kohli)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Wt. of Ref.
Standardgm.
Wt. of Sample
gm.
Measured Absor-
bance at of Ref.
Std.
Measured Absor-
bance at 268nm of
sample
% of Cefixime
Wt. of Ref.
Standardgm.
Wt. of Sample
gm
Measured Absor-
bance at of Ref.
Std.
Measured Absor-
bance at 268nm
of sample
% of Cefixime
01 0.1128 0.4019 498 502 101.75 0.1118 0.4028 502 508 101.21
02 0.1119 0.4021 492 499 101.50 0.1129 0.4031 499 505 102.14
03 0.1131 0.4012 498 488 99.34 0.1125 0.4022 492 498 102.02
Average wt of tablet 402.8 mg Averagewt.of tablet 403.6 mg
Mean 100.86% Mean 101.79%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 101.32 %Standard Deviation : ±10 %
Sarthak Biotech Pvt. Ltd. Total Page: 01-06
Assay Method Validation QC/VLD/05Name of the Product: SARMOX-500 CAPSULEPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 10/10/2007 Review Date : 09/10/2009 Revision No. : 00
PURPOSE: The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Amoxycillin in Sarmox-500 Capsule will give results with acceptable Accuracy, Linearity, Repeatibility, Relative Standard Deviation & Ruggedness when applied to the Sarmox-500 Capsule.
SCOPE : This method is applies to the procedure which is used for calculation and assay of Amoxycillin.
RESPONSIBILITY: 1. Quality Control Manager.
2. Asstt. Quality Control Chemist.
VALIDATION PARAMETERS:
1. Linearity.
2. Repeatability.
3. Relative Standard Deviation.
4. Ruggedness.
Sarthak Biotech Pvt. Ltd. Total Page: 02-06
Assay Method Validation QC/VLD/05Name of the Product: SARMOX-500 CAPSULEPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 10/10/2007 Review Date : 09/10/2009 Revision No. : 00
METHOD :
1. Weight accurately 100mg Reference standard and equivalent to 100mg. Amoxycillin as test sample. Bowl it with 20 to 30ml of distilled water and make up to produce 100ml.
2. Pipette 2ml, 3ml, 4ml, 5ml and 6ml of standard and sample solution separately in a calibrated 50ml.volumetric flask.
3. In a 50ml volumetric flask pipette 2ml Reference and test solution separately. Toeach flask add 2ml of Phosphate buffer pH 9.2 and add 1ml Vitamin B6 dye..
4. Measure the absorbance without delay at maximum at about 605nm,against respective blank and tabulate the observed data in the observation table.
5. Plot linearity curve from the different concentration corresponding area of Amoxycillin peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.
6. Plot repeatability curve from by taking Amoxycillin solution serial no. on X-axis and observed peak area on Y-axis.
7. Draw a line which should cover almost all the points. The resulted graph should be
almost parallel to the X-axis. . .
8. Calculated the result by comparison with peak area obtained in standard and sample solution respectively.
Sarthak Biotech Pvt. Ltd. Total Page: 03-06
Assay Method Validation QC/VLD/05Name of the Product: SARMOX-500 CAPSULEPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 10/10/2007 Review Date : 09/10/2009 Revision No. : 00
LINEARITY STUDY
The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range.
Linearity of assay method of Amoxycillin in Sarmox-500 Capsule shall be studied by measuring the absorbance of the standard Amoxycillin solution of 40 – 120mcg/ml concentration & range 605nm. Linearity curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR LINEARITY
Sr.No. Concentration of Amoxycillin in mcg/ml Peak area of 1. 40mcg\ml 652
2. 60mcg\ml 695
3. 80mcg\ml 728
4. 100mcg\ml 765
5. 120mcg\ml 805
Sarthak Biotech Pvt. Ltd. Total Page: 04-06Assay Method Validation QC/VLD/05Name of the Product: SARMOX-500 CAPSULEPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 10/10/2007 Review Date : 09/10/2009 Revision No. : 00
REPEATIBILITY STUDY
Repeatibility of assay method of Amoxycillin in Sarmox-500 Capsule shall be studied by measuring the absorbance of the standard Amoxycillin solution of 40mcg/ml concentration & range 605nm. Repeatibility curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR REPEATBILITY
Sr.No. Concentration in mcg\ml Peak area 1 40mcg\ml 651
2 . 40mcg\ml 653
3. 40mcg\ml 652
4. 40mcg\ml 657
5. 40mcg\ml 651
Sarthak Biotech Pvt. Ltd. Total Page: 05-06Assay Method Validation QC/VLD/05Name of the Product: SARMOX-500 CAPSULEPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 10/10/2007 Review Date : 09/10/2009 Revision No. : 00
RELATIVE STANDARD DEVIATION Test concentration of Amoxycillin used = 40mcg\ml.
Number of test solution used = 5
651+653+652+657+651 Average area = ----------------------------------- = 652.8
5
(651-652.8)2 +(653-652.8)2 +(652-652.8)2 +(657-652.8)2 +(651-652.8)2
Dev. S = ------------------------------------------------------------------------------------- 1/2
5-1
3.24+0.04+0.64+17.64+3.24 = -------------------------------------1/2
4
24.8 = -------1/2
4
= 6.2 1/2
= 2.489
2.489 x 100 RSD = ------------------ = 0.381%
652.8
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd. Total Page: 06-06
Assay Method Validation QC/VLD/05Name of the Product: SARMOX-500 CAPSULEPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 10/10/2007 Review Date : 09/10/2009 Revision No. : 00
RUGGEDNESS STUDY
The ruggedness of an analytical method is the degree of reproducibility of the test
results obtained by the analysis of the samples from same lot under a variety of normal test
conditions (i.e. different laboratories, analysts, days & Instruments).
The ruggedness of assay method of Amoxycillin in Sarmox-500 Capsule shall be
studied by analyzing the samples from same lot by two different analysts from same laboratory
or at two different laboratories by two different analysts on two different days.
RUGGEDNESS STUDY DATA
Ruggedness data for assay method by two different analysts on different days and different instrument.
Sr. No.
Analyst- I (Satpal Saini)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Analyst- II (Rajnish Kohli)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Wt. of Ref.
Standardgm.
Wt. of Sample
gm.
Measured Absor-
bance at of Ref.
Std.
Measured Absor-
bance at 605nm of
sample
% of Amoxyci
-llin
Wt. of Ref.
Standardgm.
Wt. of Sample
gm
Measured Absor-
bance at of Ref.
Std.
Measured Absor-
bance at 605nm
of sample
% of Amoxycil
-lin
01 0.1162 0.1148 655 652 100.79 0.1154 0.1168 659 655 98.57
02 0.1159 0.1162 654 658 100.38 0.1162 0.1169 652 662 101.30
03 0.1165 0.1155 653 657 101.51 0.1152 0.1162 656 650 98.60
Average fill weight 580.2 mg Average fill weight 582.2 mg
Mean 100.89% Mean 99.49%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 100.19 %Standard Deviation : ±10 %
Sarthak Biotech Pvt. Ltd. Total Page: 01-06Assay Method Validation QC/VLD/06Name of the Product: ACIGONE CAPSULEPrepared By Checked By/ Approved By
Date of Issue/Effective Date :27/09/2007 Review Date : 26/09/2009 Revision No. : 00
PURPOSE: The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Omeprazole in Acigone Capsule will give results with acceptable Accuracy, Linearity, Repeatability, Relative Standard Deviation & Ruggedness when applied to the Acigone Capsule.
SCOPE : This method is applies to the procedure which is used for calculation and assay of Omeprazole.
RESPONSIBILITY: 1. Quality Control Manager.
2. Asstt. Quality Control Chemist.
VALIDATION PARAMETERS:
1. Linearity.
2. Repeatability.
3. Relative Standard Deviation.
4. Ruggedness.
Sarthak Biotech Pvt. Ltd. Total Page: 02-06Assay Method Validation QC/VLD/06Name of the Product: ACIGONE CAPSULEPrepared By Checked By/ Approved By
Date of Issue/Effective Date :27/09/2007 Review Date : 26/09/2009 Revision No. : 00
METHOD :
1. Weight accurately 20mg Reference standard and equivalent to 20mg. Omeprazole as test sample. Dissolve it with 20 to 30ml of 0.1M Sodium Hydroxide solution shake vigorously 4-5 minutes and make up the volume up to 50ml with 0.1M NaOH solution.
2. Pipette 1ml, 2ml, 3ml, 4ml and 5ml of standard and sample solution separately in a calibrated 50ml.volumetric flask.
3. In a 50ml volumetric flask pipette 1ml Reference and test solution separately and make up the volume up to 50ml. with 0.1M Sodium Hydroxide solution.
4. Measure the absorbance without delay at maximum at about 268nm,against respective blank and tabulate the observed data in the observation table.
5. Plot linearity curve from the different concentration corresponding area of Omeprazole peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.
6. Plot repeatability curve from by taking Omeprazole solution serial no. on X-axis and observed peak area on Y-axis.
7. Draw a line which should cover almost all the points. The resulted graph should be almost parallel to the X-axis. . .
8. Calculated the result by comparison with peak area obtained in standard and sample solution respectively.
Sarthak Biotech Pvt. Ltd. Total Page: 03-06Assay Method Validation QC/VLD/06Name of the Product: ACIGONE CAPSULEPrepared By Checked By/ Approved By
Date of Issue/Effective Date :27/09/2007 Review Date : 26/09/2009 Revision No. : 00
LINEARITY STUDY
The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range.
Linearity of assay method of Omeprazole in Acigone Capsule shall be studied by measuring the absorbance of the standard Omeprazole solution of 8 – 40mcg/ml concentration & range 268nm. Linearity curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR LINEARITY
Sr.No. Concentration of Amoxycillin in mcg\ml Peak area of 1. 8mcg\ml 397
2. 16mcg\ml 421
3. 24mcg\ml 495
4. 32mcg\ml 565
5. 40mcg\ml 605
Sarthak Biotech Pvt. Ltd. Total Page: 04-06Assay Method Validation QC/VLD/06Name of the Product: ACIGONE CAPSULEPrepared By Checked By/ Approved By
Date of Issue/Effective Date :27/09/2007 Review Date : 26/09/2009 Revision No. : 00
REPEATIBILITY STUDY
Repeatibility of assay method of Omeprazole in Acigone Capsule shall be studied by measuring the absorbance of the standard Omeprazole solution of 8 mcg/ml concentration & range 268nm. Repeatibility curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR REPEATIBILITY
Sr.No. Concentration in mcg\ml Peak area 1 8mcg\ml 398
2 . 8mcg\ml 401
3. 8mcg\ml 397
4. 8mcg\ml 399
5. 8mcg\ml 398
Sarthak Biotech Pvt. Ltd. Total Page: 05-06Assay Method Validation QC/VLD/06Name of the Product: ACIGONE CAPSULEPrepared By Checked By/ Approved By
Date of Issue/Effective Date :27/09/2007 Review Date : 26/09/2009 Revision No. : 00
RELATIVE STANDARD DEVIATION Test concentration of Amoxycillin used = 8mcg\ml. Number of test solution used = 5
398+401+397+399+398 Average area = ----------------------------------- = 398.6
5
(398-398.6)2 +(401-398.6)2 +(397-398.6)2 +(399-398.6)2 +(398-398.6)2
Dev. S = --------------------------------------------------------------------------------------- 1/2
5-1
0.36+5.76 +2.56+0.16+0.36 = ---------------------------------------1/2
4
9.2 = -------1/2
4
= 2.3 1/2
= 1.5165
1.5165 x 100 RSD = ------------------ = 0.3804%
398.6
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd. Total Page: 06-06Assay Method Validation QC/VLD/06Name of the Product: ACIGONE CAPSULEPrepared By Checked By/ Approved By
Date of Issue/Effective Date :27/09/2007 Review Date : 26/09/2009 Revision No. : 00
RUGGEDNESS STUDY
The ruggedness of an analytical method is the degree of reproducibility of the test
results obtained by the analysis of the samples from same lot under a variety of normal test
conditions (i.e. different laboratories, analysts, days & Instruments).
The ruggedness of assay method of Omeprazole in Acigone Capsule shall be studied
by analyzing the samples from same lot by two different analysts from same laboratory or at
two different laboratories by two different analysts on two different days.
RUGGEDNESS STUDY DATA
Ruggedness data for assay method by two different analysts on different days and different instrument.
Sr. No.
Analyst- I (Satpal Saini)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Analyst- II (Rajnish Kohli)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Wt. of Ref.
Standard gm.
Wt. of Sample
gm.
Measured Absor-
bance at of Ref.
Std.
Measured Absor-
bance at 268nm of
sample
% of Omepraz
-ole
Wt. of Ref.
Standard gm.
Wt. of Sample
gm
Measured Absor-
bance at of Ref.
Std.
Measured Absor-
bance at 268nm
of sample
% of Omepra-
zole
01 0.2657 0.2940 395 399 100.40 0.2672 0.2926 403 399 99.48
02 0.2673 0.2927 401 396 99.20 0.2657 0.2961 398 401 99.47
03 0.2661 0.2939 393 396 100.38 0.2673 0.2926 396 393 99.75
Average fill weight 293.4 mg Average fill weight 293.4 mg
Mean 99.99% Mean 99.57%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 99.78
Standard Deviation : ±10%
Sarthak Biotech Pvt. Ltd. Total Page: 01-06Assay Method Validation QC/VLD/07Name of the Product: SARMOX DRY SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 29/10/2007 Review Date : 28/10/2009 Revision No. : 00
PURPOSE: The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Amoxycillin in Sarmox Dry Syrup will give results with acceptable Accuracy, Linearity, Repeatability, Relative Standard Deviation & Ruggedness when applied to the Sarmox Dry Syrup.
SCOPE : This method is applies to the procedure which is used for calculation and assay of Amoxycillin.
RESPONSIBILITY: 1. Quality Control Manager.
2. Asstt. Quality Control Chemist.
VALIDATION PARAMETERS:
1. Linearity.
2. Repeatibility.
3. Relative Standard Deviation.
4. Ruggedness.
Sarthak Biotech Pvt. Ltd. Total Page: 02-06Assay Method Validation QC/VLD/07Name of the Product: SARMOX DRY SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 29/10/2007 Review Date : 28/10/2009 Revision No. : 00
METHOD :
1. Weight accurately 100mg Reference standard and equivalent to 100mg. Amoxycillin as test sample. Bowl it with 20 to 30ml of distilled water and make up to produce 100ml.
2. Pipette 2ml, 4ml, 5ml, 6ml and 7ml of standard and sample solution separately in a calibrated 50ml.volumetric flask.
3. In a 50ml volumetric flask pipette 2ml Reference and test solution separately. To each flask add 2ml of Phosphate buffer pH 9.2 and add 1ml Vitamin B6 dye. And make up the volume up to 50ml with water.
4. Measure the absorbance without delay at maximum at about 605nm,against respective blank and tabulate the observed data in the observation table.
5. Plot linearity curve from the different concentration corresponding area of Amoxycillin peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.
6. Plot repeatability curve from by taking Amoxycillin solution serial no. on X-axis and observed peak area on Y-axis.
7. Draw a line which should cover almost all the points. The resulted graph should be
almost parallel to the X-axis. . .
8. Calculated the result by comparison with peak area obtained in standard and sample solution respectively.
Sarthak Biotech Pvt. Ltd. Total Page: 03-06Assay Method Validation QC/VLD/07Name of the Product: SARMOX DRY SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 29/10/2007 Review Date : 28/10/2009 Revision No. : 00
LINEARITY STUDY
The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range.
Linearity of assay method of Amoxycillin in Sarmox Dry Syrup shall be studied by measuring the absorbance of the standard Amoxycillin in solution of 40 – 120mcg/ml concentration & range 605nm. Linearity curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR LINEARITY
Sr.No. Concentration of Amoxycillin in mcg\ml Peak area of 1. 40mcg\ml 651
2. 60mcg\ml 695
3. 80mcg\ml 726
4. 100mcg\ml 775
5. 120mcg\ml 804
Sarthak Biotech Pvt. Ltd. Total Page: 04-06Assay Method Validation QC/VLD/07Name of the Product: SARMOX DRY SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 29/10/2007 Review Date : 28/10/2009 Revision No. : 00
REPEATIBILITY STUDY
Repeatibility of assay method of Amoxycillin in Sarmox Dry Syrup shall be studied by measuring the absorbance of the standard Amoxycillin in solution of 40 mcg/ml concentration & range 605nm. Repeatibility curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR REPEATIBILITY
Sr.No. Concentration in mcg\ml Peak area 1 40mcg\ml 651
2 . 40mcg\ml 652
3. 40mcg\ml 657
4. 40mcg\ml 660
5. 40mcg\ml 655
Sarthak Biotech Pvt. Ltd. Total Page: 05-06Assay Method Validation QC/VLD/07Name of the Product: SARMOX DRY SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 29/10/2007 Review Date : 28/10/2009 Revision No. : 00
RELATIVE STANDARD DEVIATION
Test concentration of Amoxycillin used = 40mcg\ml. Number of test solution used = 5
651+652+657+660+655 Average area = ----------------------------------- = 655
5
(651-655)2 +(652-655)2 +(657-655)2 +(660-655)2 +(655-655)2
Dev. S = ------------------------------------------------------------------------------ 1/2
5-1
16+ 9+ 4 + 25 + 0 = ---------------------------------------1/2
4
54 = -------1/2
4
= 13.5 1/2
= 3.6742
3.6742 x 100 RSD = ------------------ = 0.560%
655 Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd. Total Page: 06-06Assay Method Validation QC/VLD/07Name of the Product: SARMOX DRY SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 29/10/2007 Review Date : 28/10/2009 Revision No. : 00
RUGGEDNESS STUDY
The ruggedness of an analytical method is the degree of reproducibility of the test
results obtained by the analysis of the samples from same lot under a variety of normal test
conditions (i.e. different laboratories, analysts, days & Instruments).
The ruggedness of assay method of Amoxycillin in Sarmox Dry Syrup shall be studied
by analyzing the samples from same lot by two different analysts from same laboratory or at
two different laboratories by two different analysts on two different days.
RUGGEDNESS STUDY DATA
Ruggedness data for assay method by two different analysts on different days and different instrument.
Sr. No.
Analyst- I (Satpal Saini)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Analyst- II (Rajnish Kohli)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Wt. of Ref.
Standardgm.
Wt. of Sample
ml
Measured
Absor-bance
at of Ref.
Std.
Measured Absor-
bance at 605nm of
sample
% of Amoxy-
cillin
Wt. of Ref.
Standardgm.
Wt. of Sample
ml
Measured Absor-
bance at of Ref.
Std.
Measured Absor-
bance at 605nm
of sample
% of Amoxy-
cillin
01 0.1161 4ml 653 656 100.54 0.1159 4ml 657 652 99.15
02 0.1168 4ml 658 665 101.76 0.1162 4ml 659 663 100.18
03 0.1155 4ml 652 656 100.82 0.1155 4ml 657 654 99.11
Label Claim 125mg Amoxycillin/ 5ml Label Claim 125mg Amoxycillin/ 5ml
Mean 100.82% Mean 100.01%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 100.41 %Standard Deviation : ±10 %
Sarthak Biotech Pvt. Ltd. Total Page: 01-06Assay Method Validation QC/VLD/08Name of the Product: CEFSAR-125 DRY SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 17/01/2008 Review Date : 16/01/2010 Revision No. : 00
PURPOSE : The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Cephalexin in Cefasar-125 Dry Syrup will give results with acceptable Accuracy, Linearity, Repeatibility, Relative Standard Deviation & Ruggedness when applied to the Cefasar-125 Dry Syrup.
SCOPE : This method is applies to the procedure which is used for calculation and assay
of Cephalexin.
RESPONSIBILITY: 1. Quality Control Manager.
2. Asstt. Quality Control Chemist.
VALIDATION PARAMETERS:
1. Linearity;
2. Repeatibility.
3. Relative Standard Deviation.
4. Ruggedness.
Sarthak Biotech Pvt. Ltd. Total Page: 02-06Assay Method Validation QC/VLD/08Name of the Product: CEFSAR-125 DRY SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 17/01/2008 Review Date : 16/01/2010 Revision No. : 00
METHOD :
1. Weight accurately equivalent 250mg of Cephalexin and dissolve in 20ml of Neutral Methanol and titrate with 0.1M Sodium Hydroxide using Bromomethyl blue as indicator, perform a blank for necessary correction. Each ml. of 0.1M Sodium Hydroxide is equivalent to 0.03474gm. of Cephalexin.
2. Weight 250mg, 300mg, 350mg., 400mg. & 450mg. Cephalexin in 20ml. of Neutral
Methanol & titrate with 0.1M NaOH using Bromomethyl blue as indicator. Each ml. of 0.1m Sodium Hydroxide is equivalent to 0.03474gm. of Cephalexin.
3. Note the volume consumed & tabulate the observed data in observation table.
4. Plot linearity curve from the different concentration corresponding area of Cephalexin peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.
5. Plot repeatability curve from by taking Cephalexin solution serial no. on X-axis and observed peak area on Y-axis.
6. Draw a line which should cover almost all the points. The resulted graph should be
almost parallel to the X-axis. . .
7. Calculated the result by calculation volume consumed by the Cephalexin.
Sarthak Biotech Pvt. Ltd. Total Page: 03-06Assay Method Validation QC/VLD/08Name of the Product: CEFSAR-125 DRY SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 17/01/2008 Review Date : 16/01/2010 Revision No. : 00
LINEARITY STUDY
The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range.
Linearity of assay method of Cephalexin in Cefsar-125 Dry Syrup shall be studied by 250mg-to 450 mg Cephalexin.
OBSERVATION TABLE FOR LINEARITY
( Molarity of 0.1M NaOH is 0.102305)
Sr.No. Weight of Cephalexin in mg Volume consumed 1. 250mg. 7ml
2. 300mg. 8.6ml
3. 350mg. 9.7ml
4. 400mg. 11.2ml
5. 450mg. 12.7ml
Sarthak Biotech Pvt. Ltd. Total Page: 04-06Assay Method Validation QC/VLD/08Name of the Product: CEFSAR-125 DRY SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 17/01/2008 Review Date : 16/01/2010 Revision No. : 00
REPEATIBILITY STUDY
Repeatibility of assay method of Cephalexin in Cefsar-125 Dry Syrup shall be studied by 250 mg Cephalexin.
OBSERVATION TABLE FOR REPEATIBILITY
( Molarity of 0.1M NaOH is 0.102305)
Sr.No. Weight of Cephalexin Volume consumed
1 250mg. 7ml
2 . 250mg. 6.9ml
3. 250mg. 7.1ml
4. 250mg. 7.2ml
5. 250mg. 6.9ml
Sarthak Biotech Pvt. Ltd. Total Page: 05-06Assay Method Validation QC/VLD/08Name of the Product: CEFSAR-125 DRY SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 17/01/2008 Review Date : 16/01/2010 Revision No. : 00
RELATIVE STANDARD DEVIATION
Test concentration of Cephalexin used = 250mg. Number of test solution used = 5 7+6.9+7.1+7.2+6.9 Average volume = ----------------------------------- = 7.02
5
(7- 7.02)2 +(6.9-7.02)2 +(7.1-7.02)2 +(7.2-7.02)2 +(6.9-7.02)2
Dev. S = --------------------------------------------------------------------------------------- 1/2
5-1
0.0004+0.0144+0.0064+0.0324+0.0144 = ------------------------------------------------------1/2
4
0.068 = ------- 1/2
4
= 0.017 1/2
= 0.130
0.130 x 100 RSD = ------------------ = 1.857%
7.02
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd. Total Page: 06-06Assay Method Validation QC/VLD/08Name of the Product: CEFSAR-125 DRY SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 17/01/2008 Review Date : 16/01/2010 Revision No. : 00
RUGGEDNESS STUDY
The ruggedness of an analytical method is the degree of reproducibility of the test
results obtained by the analysis of the samples from same lot under a variety of normal test
conditions (i.e. different laboratories, analysts, days & Instruments).
The ruggedness of assay method of Cephalexin inCefsar-125 Dry Syrup shall be
studied by analyzing the samples from same lot by two different analysts from same laboratory
or at two different laboratories by two different analysts on two different days.
RUGGEDNESS STUDY DATA
Ruggedness data for assay method by two different analysts on different days and different instrument.
Sr. No.
Analyst- I (Satpal Saini)Date of analysis :Instrument & Model : Burette 50ml (Borosil)Location : Chemical Lab.
Analyst- II (Rajnish Kohli)Date of analysis :Instrument & Model : Burette 50ml (Borosil)Location : Chemical Lab.
Wt. of Sample
Molarity of 0.1M NaOH
Final Volume
used
Assay Wt. of Sample
Molarity of 0.1M NaOH
Final Volume used
Assay
01 10ml 0.102305 7.0ml 99.51 10ml 0.102305 7.1ml 100.94
02 10ml 0.102305 6.9ml 98.09 10ml 0.102305 7.0ml 99.51
03 10ml 0.102305 7.0ml 99.51 10ml 0.102305 7.0ml 99.52
Label Claim 125mg Cephalexin/5ml Label Claim 125mg Cephalexin/5ml
Mean 99.04% Mean 99.99%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 99.52%Standard Deviation : ±10%
Sarthak Biotech Pvt. Ltd. Total Page: 01-05Assay Method Validation QC/VLD/09Name of the Product: CHLORTEX MOUTH WASHPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 10/01/2008 Review Date : 09/01/2010 Revision No. : 00
PURPOSE : The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Chlorhexidine Gluconate in Chlortex Mouth Wash will give results with acceptable Accuracy, Linearity, Repeatability, Relative Standard Deviation & Ruggedness when applied to the Cefasar-125 Dry Syrup.
SCOPE : This method is applies to the procedure which is used for calculation and assay of Chlorhexidine Gluconate.
RESPONSIBILITY: 1. Quality Control Manager.
2. Asstt. Quality Control Chemist.
VALIDATION PARAMETERS:
1. Linearity.
2. Repeatability.
3. Relative Standard Deviation.
4. Ruggedness.
Sarthak Biotech Pvt. Ltd. Total Page: 02-05Assay Method Validation QC/VLD/09Name of the Product: CHLORTEX MOUTH WASHPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 10/01/2008 Review Date : 09/01/2010 Revision No. : 00
METHOD :
1. Weigh accurately equivalent of 200mg. of Chlorhexidine Gluconate Reference standard and dissolve in 100ml of distill water.
2. Further pipette 5ml, 10ml, 15ml, 20ml & 25ml of standard and dilute to 100ml. with water.
3. Weigh accurately equivalent to 5mg., 10mg., 15mg., 20mg. & 25mg. of sample, basified with 10M NaOH and extracted with Chloroform (2 x 20ml). Combine Chloroform layer and dilute to 50ml with water. Further prepare dilution of 50mcg / ml. with water.
4. To 1 ml. of each above standard and sample dilutions add 3ml of dye solution and 5ml. of Chloroform. Shake well for 3 to 4 minutes.
5. Measure the absorbance without delay at maximum at about 410nm, against respective blank and tabulate the observed data in the observation table.
6. Plot linearity curve from the different concentration corresponding area of Chlorhexidine Gluconate peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.
7. Plot repeatability curve from by taking Chlorhexidine Gluconate solution serial no. on X-axis and observed peak area on Y-axis.
8. Draw a line which should cover almost all the points. The resulted graph should be
almost parallel to the X-axis. . .
9. Calculated the result by comparison with peak area obtained in standard and sample solution respectively.
Sarthak Biotech Pvt. Ltd. Total Page: 03-05Assay Method Validation QC/VLD/09Name of the Product: CHLORTEX MOUTH WASHPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 10/01/2008 Review Date : 09/01/2010 Revision No. : 00
LINEARITY STUDY
The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range.
Linearity of assay method of Chlorhexidine Gluconate in Chlortex Mouthwash shall be studied by measuring the absorbance of the standard Chlorhexidine Gluconate in solution of 100 – 500mcg/ml concentration & range 410nm. Linearity curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR LINEARITY
Sr.No. Concentration of Chlorhexidine Gluconate in mcg\ml Peak area of 1. 100mcg\ml 546
2. 200mcg\ml 642
3. 300mcg\ml 726
4. 400mcg\ml 791
5. 500mcg\ml 805
Sarthak Biotech Pvt. Ltd. Total Page: 04-06Assay Method Validation QC/VLD/09Name of the Product: CHLORTEX MOUTH WASHPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 10/01/2008 Review Date : 09/01/2010 Revision No. : 00
REPEATIBILITY STUDY
Repeatibility of assay method of Chlorhexidine Gluconate in Chlortex Mouthwash shall be studied by measuring the absorbance of the standard Chlorhexidine Gluconate in solution of 100 mcg/ml concentration & range 410nm. Repeatibility curve shall be plotted for absorbance against concentration
OBSERVATION TABLE FOR REPEATIBILITY
Sr.No. Concentration in mcg\ml Peak area
1 100mcg\ml 544
2 . 100mcg\ml 545
3. 100mcg\ml 548
4. 100mcg\ml 549
5. 100mcg\ml 546
Sarthak Biotech Pvt. Ltd. Total Page: 05-06Assay Method Validation QC/VLD/09Name of the Product: CHLORTEX MOUTH WASHPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 10/01/2008 Review Date : 09/01/2010 Revision No. : 00
RELATIVE STANDARD DEVIATION
Test concentration of Chlorhexidine Gluconate used = 100mcg\ml. Number of test solution used = 5 544+545+548+549+546 Average area = ----------------------------------- = 546.4
5
(544-546.4)2 +(545-546.4)2 +(548-546.4)2 +(549-546.4)2 +(546-546.4)2
Dev. S = --------------------------------------------------------------------------------------- 1/2
5-1
5.76+2.005+3.2 +1.99 +0.8 = ---------------------------------------1/2
4 13.755 = ----- ----1/2
4 = 3.439 1/2
= 1.854
1.854 x 100 RSD = ------------------ = 0.339 %
546.4
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd. Total Page: 06-06Assay Method Validation QC/VLD/09Name of the Product: CHLORTEX MOUTH WASHPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 10/01/2008 Review Date : 09/01/2010 Revision No. : 00
RUGGEDNESS STUDY
The ruggedness of an analytical method is the degree of reproducibility of the test
results obtained by the analysis of the samples from same lot under a variety of normal test
conditions (i.e. different laboratories, analysts, days & Instruments).
The ruggedness of assay method of Chlorhexidine Gluconate in Chlortex Mouthwash
shall be studied by analyzing the samples from same lot by two different analysts from same
laboratory or at two different laboratories by two different analysts on two different days.
RUGGEDNESS STUDY DATA
Ruggedness data for assay method by two different analysts on different days and different instrument.
Sr. No.
Analyst- I (Rajnish Kohli)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Analyst- II (Satpal Saini) Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Wt. of Ref.
Standard gm.
Wt. of Sample
ml
Measured
Absor-bance
at of Ref.
Std.
Measured Absor-
bance at 410nm of
sample
% of Chlorhexi
dine Gluconate
Wt. of Ref.
Standard gm.
Wt. of Sample
ml
Measured Absor-
bance at of Ref.
Std.
Measured
Absor-bance at 410nm
of sample
% of Chlorhexid
ine Gluconate
01 0.4998 50ml 544 545 100.49 0.5090 50ml 548 547 99.99
02 0.4989 50ml 546 542 99.04 0.4992 50ml 542 545 100.39
03 0.5012 50ml 543 545 99.69 0.4984 50ml 542 545 100.23
Label Claim 0.2% Chlorhexidine
Gluconate
Label Claim 0.2% Chlorhexidine
Gluconate
Mean 99.74% Mean 100.02%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 99.98 %Standard Deviation : ±10 %
Sarthak Biotech Pvt. Ltd. Total Page: 01-16Assay Method Validation QC/VLD/10Name of the Product: BRONSYP SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 24/01/2008 Review Date : 23/01/2010 Revision No. : 00
PURPOSE : The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Salbutamol Sulphate, Bromhexine Hcl & Guaiphensin in Bronsyp Syrup will give results with acceptable Accuracy, Linearity, Repeatability, Relative Standard Deviation & Ruggedness when applied to the Bronsyp Syrup.
SCOPE : This method is applies to the procedure which is used for calculation and assay of Salbutamol Sulphate, Bromhexine Hcl & Guaiphensin.
RESPONSIBILITY: 1. Quality Control Manager.
2. Asstt. Quality Control Chemist.
VALIDATION PARAMETERS:
1. Linearity.
2. Repeatability.
3. Relative Standard Deviation.
4. Ruggedness.
Sarthak Biotech Pvt. Ltd. Total Page: 02-16Assay Method Validation QC/VLD/10Name of the Product: BRONSYP SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 24/01/2008 Review Date : 23/01/2010 Revision No. : 00
METHOD : (Salbutamol Sulphate)
1. Weigh accurately 100mg. of Salbutamol Sulphate as Reference standard and dissolve in 100 ml of water and prepare the dilution 100mcg/ml. in water.
2. Accurately weigh quantity of sample is extracted with water to get 100mcg./ml in
respect of Salbutamol Sulphate.
3. Pipette 3ml, 4ml, 5ml, 6ml & 7ml of standard and sample solution and add 3ml of Sodium Carbonate solution, 0.2ml of P-aminophenol reagent and volume is adjusted to 10ml with water.
4. Measure the absorbance without delay at maximum at about 635nm, against respective
blank and tabulate the observed data in the observation table.
5. Plot linearity curve from the different concentration corresponding area of Salbutamol Sulphate peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.
6. Plot repeatability curve from by taking Salbutamol Sulphate solution serial no. on X-axis and observed peak area on Y-axis.
7. Draw a line which should cover almost all the points. The resulted graph should be
almost parallel to the X-axis. . .
8. Calculated the result by comparison with peak area obtained in standard and sample solution respectively.
Sarthak Biotech Pvt. Ltd. Total Page: 03-16Assay Method Validation QC/VLD/10Name of the Product: BRONSYP SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 24/01/2008 Review Date : 23/01/2010 Revision No. : 00
LINEARITY STUDY
The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range.
Linearity of assay method of Salbutamol Sulphate in Bronsyp Syrup shall be studied by measuring the absorbance of the standard Salbutamol Sulphate in solution of 30 – 70mcg/ml concentration & range 635nm. Linearity curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR LINEARITY
Sr.No. Concentration of Salbutamol Sulphate in mcg\ml Peak area of 1. 30mcg\ml 695
2. 40mcg\ml 739
3. 50mcg\ml 795
4. 60mcg\ml 829
5. 70mcg\ml 888
Sarthak Biotech Pvt. Ltd. Total Page: 04-16Assay Method Validation QC/VLD/10Name of the Product: BRONSYP SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 24/01/2008 Review Date : 23/01/2010 Revision No. : 00
REPEATIBILITY STUDY
Repeatibility of assay method of Salbutamol Sulphate in Bronsyp Syrup shall be
studied by measuring the absorbance of the standard Salbutamol Sulphate in solution of 30
mcg/ml concentration & range 635nm. Repeatibility curve shall be plotted for absorbance
against concentration.
OBSERVATION TABLE FOR REPEATIBILITY
S.No. Concentration in mcg\ml Peak area
1 30mcg\ml 698
2 . 30mcg\ml 700
3. 30mcg\ml 697
4. 30mcg\ml 695
5. 30mcg\ml 703
Sarthak Biotech Pvt. Ltd. Total Page: 05-16Assay Method Validation QC/VLD/10Name of the Product: BRONSYP SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 24/01/2008 Review Date : 23/01/2010 Revision No. : 00
RELATIVE STANDARD DEVIATION
Test concentration of Salbutamol Sulphate used = 30mcg\ml. Number of test solution used = 5 698+700+697+695+703 Average area = ----------------------------------- = 698.6
5
(698-698.6)2 +(700-698.6)2 +(697-698.6)2 +(695-698.6)2 +(703-698.6)2
Dev. S = --------------------------------------------------------------------------------------- 1/2
5-1
0.36+ 1.96+ 2.56+ 12.96 + 19.36 = -------------------------------------------1/2
4 37.2 = ----- ----1/2
4
= 9.3 1/2
= 3.049
3.049 x 100 RSD = ------------------ = 0.436 %
698.6
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd. Total Page: 06-16Assay Method Validation QC/VLD/10Name of the Product: BRONSYP SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 24/01/2008 Review Date : 23/01/2010 Revision No. : 00
RUGGEDNESS STUDY
The ruggedness of an analytical method is the degree of reproducibility of the test
results obtained by the analysis of the samples from same lot under a variety of normal test
conditions (i.e. different laboratories, analysts, days & Instruments).
The ruggedness of assay method of Salbutamol Sulphate in Bronsyp Syrup shall be
studied by analyzing the samples from same lot by two different analysts from same laboratory
or at two different laboratories by two different analysts on two different days.
RUGGEDNESS STUDY DATA
Ruggedness data for assay method by two different analysts on different days and different instrument.
Sr. No.
Analyst- I (Rajnish Kohli)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Analyst- II (Satpal Saini)Date of analysis : Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Wt. of Ref.
Standardgm.
Wt. of Sample
ml
Measured
Absor-bance
at of Ref.
Std.
Measured Absor-
bance at 635nm of
sample
% of Salbutam
ol
Wt. of Ref.
Standardgm.
Wt. of Sample
ml
Measured Absor-
bance at of Ref.
Std.
Measured Absor-
bance at 635nm
of sample
% of Salbutam
ol
01 0.1209 5ml 698 702 100.32 0.1218 5ml 695 694 100.35
02 0.1216 5ml 694 697 100.76 0.1206 5ml 694 694 99.22
03 0.1206 5ml 695 698 99.77 0.1218 5ml 698 701 100.93
Label Claim 2mg. / 5ml Label Claim 2 mg. / 5ml
Mean 100.28% Mean 100.93%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 100.16%Standard Deviation : ±10 %
Sarthak Biotech Pvt. Ltd. Total Page: 07-16Assay Method Validation QC/VLD/10Name of the Product: BRONSYP SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 24/01/2008 Review Date : 23/01/2010 Revision No. : 00
METHOD : (Bromhexine Hcl)
1. Weigh accurately 40mg. of Bromhexine Hcl. and transfer to a separating funnel, basify with 5ml of dilute Ammonia solution , add 25ml. of water and extract with for 25ml portions of Chloroform layer through anhydrous Sodium Sulphate previously washed with Chloroform. Evaporate Chloroform layer and dissolve residue in 0.1M Methanolic Hydrochloric Acid to produce 100ml. (400mcg./ ml.). Further make the dilution with Methanolic Hydrochloric Acid to get final concentration of 40mcg/ml.
2. Measure accurately sample equivalent to 4mg. of Bromhexine Hcl., add 25ml. water
and continue with the procedure as described under preparation of standard solution.
3. Pipette 2ml, 3ml, 4ml, 5ml & 6ml of standard and sample solution, add 8ml of P-dimethyl-aminobenzaldehyde (10%w/v in Methanol) & 5ml. of Hydrochloric Acid mix and allow the stand for 10 minutes and then make up the volume to 25ml. with water.
4. Measure the absorbance at maximum at about 425nm, against respective blank and
tabulate the observed data in the observation table.
5. Plot linearity curve from the different concentration corresponding area of Bromhexine Hcl peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.
6. Plot repeatability curve from by taking Bromhexine Hcl. solution serial no. on X-axis and observed peak area on Y-axis.
7. Draw a line which should cover almost all the points. The resulted graph should be
almost parallel to the X-axis. . .
8. Calculated the result by comparison with peak area obtained in standard and sample solution respectively.
Sarthak Biotech Pvt. Ltd. Total Page: 08-16Assay Method Validation QC/VLD/10Name of the Product: BRONSYP SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 24/01/2008 Review Date : 23/01/2010 Revision No. : 00
LINEARITY STUDY
The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range.
Linearity of assay method of Bromhexine Hcl in Bronsyp Syrup shall be studied by measuring the absorbance of the standard Bromhexine Hcl in solution of 3.2 – 9.6mcg/ml concentration & range 425nm. Linearity curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR LINEARITY
Sr.No. Concentration of Bromhexine Hcl. in mcg\ml Peak area of
1. 3.2mcg\ml 545
2. 4.8mcg\ml 598
3. 6.4mcg\ml 642
4. 8.0mcg\ml 691
5. 9.6mcg\ml 732
Sarthak Biotech Pvt. Ltd. Total Page: 09-16Assay Method Validation QC/VLD/10Name of the Product: BRONSYP SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 24/01/2008 Review Date : 23/01/2010 Revision No. : 00
REPEATIBILITY STUDY
Repeatibility of assay method of Bromhexine Hcl in Bronsyp Syrup shall be studied by measuring the absorbance of the standard Bromhexine Hcl in solution of 3.2mcg/ml concentration & range 425nm. Repeatibility curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR REPEATIBILITY
Sr. No. Concentration in mcg\ml Peak area
1 3.2mcg\ml 545
2 . 3.2mcg\ml 547
3. 3.2mcg\ml 548
4. 3.2mcg\ml 545
5. 3.2mcg\ml 542
Sarthak Biotech Pvt. Ltd. Total Page: 10-16Assay Method Validation QC/VLD/10Name of the Product: BRONSYP SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 24/01/2008 Review Date : 23/01/2010 Revision No. : 00
RELATIVE STANDARD DEVIATION Test concentration of Bromhexine Hcl. used = 3.2mcg\ml. Number of test solution used = 5
545+547+548+545+542 Average area = ----------------------------------- = 545.4
5
(545-545.4)2 +(547-545.4 )2 +(548-545.4)2 +(545-545.4)2 +(542-545.4)2
Dev. S = --------------------------------------------------------------------------------------- 1/2
5-1
0.16+2.56+6.76+0.16+11.56 = --------------------------------------------1/2
4 21.2 = ----- ----1/2
4
= 5.3 1/2
= 2.302
2.302 x 100 RSD = ------------------ = 0.422 %
545.4
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd. Total Page: 11-16Assay Method Validation QC/VLD/10Name of the Product: BRONSYP SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 24/01/2008 Review Date : 23/01/2010 Revision No. : 00
RUGGEDNESS STUDY
The ruggedness of an analytical method is the degree of reproducibility of the test
results obtained by the analysis of the samples from same lot under a variety of normal test
conditions (i.e. different laboratories, analysts, days & Instruments).
The ruggedness of assay method of Bromhexine Hcl in Bronsyp Syrup shall be
studied by analyzing the samples from same lot by two different analysts from same laboratory
or at two different laboratories by two different analysts on two different days.
RUGGEDNESS STUDY DATA
Ruggedness data for assay method by two different analysts on different days and different instrument.
Sr. No.
Analyst- I (Satpal Saini)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Analyst- II (Rajnish Kohli)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Wt. of Ref.
Standardgm.
Wt. of Sample
ml
Measured
Absor-bance
at of Ref.
Std.
Measured Absor-
bance at 425nm of
sample
% of Bromhex
ine
Wt. of Ref.
Standardgm.
Wt. of Sample
ml
Measured Absor-
bance at of Ref.
Std.
Measured Absor-
bance at 425nm
of sample
% of Bromhexi
ne
01 0.0398 5ml 548 546 99.13 0.0396 5ml 545 542 98.45
02 0.0396 5ml 544 548 99.72 0.0399 5ml 541 543 100.11
03 0.0399 5ml 543 545 100.11 0.0401 5ml 542 543 100.43
Label Claim 4mg. Bromhexine/5ml Label Claim 4mg. Bromhexine/5ml
Mean 99.65% Mean 99.66%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 99.66Standard Deviation : ±10%
Sarthak Biotech Pvt. Ltd. Total Page: 12-16Assay Method Validation QC/VLD/10Name of the Product: BRONSYP SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 24/01/2008 Review Date : 23/01/2010 Revision No. : 00
METHOD : (Guaiphenesin)
1. Weigh accurately 100mg. of Guaiphenesin, transfer to a separating funnel. Add 20ml of water and 5ml. of Sodium hydroxide solution (20%w/v). Extract with for 40ml portions of petroleum ether (60-80oC) . Discard ether layer, collect the aqueous layer and extract for 20ml portions of chloroform. Pass combined chloroform layer throught anhydrous sodium Sulphate and make up to 100ml with chloroform.
2. Measure accurately sample equivalent to 100mg. of substance, add 20ml of water and
continue the procedure as described under preparation of standard solution.
3. Pipette 5ml, 6ml, 7ml, 8ml & 10ml of standard and sample solution, and make up the volume 100ml. with methanol.
4. Measure the absorbance at maximum at about 275nm, against respective blank and tabulate the observed data in the observation table.
5. Plot linearity curve from the different concentration corresponding area of Guaiphenesin peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.
6. Plot repeatability curve from by taking Guaiphenesin solution serial no. on X-axis and observed peak area on Y-axis.
7. Draw a line which should cover almost all the points. The resulted graph should be
almost parallel to the X-axis. . .
8. Calculated the result by comparison with peak area obtained in standard and sample solution respectively.
Sarthak Biotech Pvt. Ltd. Total Page: 13-16Assay Method Validation QC/VLD/10Name of the Product: BRONSYP SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 24/01/2008 Review Date : 23/01/2010 Revision No. : 00
LINEARITY STUDY
The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range.
Linearity of assay method of Guaiphenesin in Bronsyp Syrup shall be studied by measuring the absorbance of the standard Guaiphenesin in solution of 50 – 100mcg/ml concentration & range 275nm. Linearity curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR LINEARITY
Sr.No. Concentration of Guaiphenesin in mcg\ml Peak area of
1. 50mcg\ml 419
2. 60mcg\ml 460
3. 70mcg\ml 502
4. 80mcg\ml 542
5. 100mcg\ml 605
Sarthak Biotech Pvt. Ltd. Total Page: 14-16Assay Method Validation QC/VLD/10Name of the Product: BRONSYP SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 24/01/2008 Review Date : 23/01/2010 Revision No. : 00
REPEATIBILITY STUDY
Repeatibility of assay method of Guaiphenesin in Bronsyp Syrup shall be studied by measuring the absorbance of the standard Guaiphenesin in solution of 50 mcg/ml concentration & range 275nm. Repeatibility curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR REPEATIBILITY
Sr.No. Concentration in mcg\ml Peak area
1 50mcg\ml 421
2 . 50mcg\ml 419
3. 50mcg\ml 424
4. 50mcg\ml 422
5. 50mcg\ml 420
Sarthak Biotech Pvt. Ltd. Total Page: 15-16
Assay Method Validation QC/VLD/10Name of the Product: BRONSYP SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 24/01/2008 Review Date : 23/01/2010 Revision No. : 00
RELATIVE STANDARD DEVIATION
Test concentration of Guaiphensin used = 50mcg\ml. Number of test solution used = 5
421+419+424+422+420 Average area = ----------------------------------- = 421.2
5
(421-421.2)2 +(419-421.2)2 +(424-421.2)2 +(422-421.2)2 +(420-421.2)2
Dev. S = --------------------------------------------------------------------------------------- 1/2
5-1
0.04+4.84+ 7.84 + 0.64 + 1.44 = --------------------------------------------1/2
4 14.8 = ----- ----1/2
4
= 3.7 1/2
= 1.924
1.924 x 100 RSD = ------------------ = 0.457 %
421.2
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd. Total Page: 16-16Assay Method Validation QC/VLD/10Name of the Product: BRONSYP SYRUPPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 24/01/2008 Review Date : 23/01/2010 Revision No. : 00
RUGGEDNESS STUDYThe ruggedness of an analytical method is the degree of reproducibility of the test
results obtained by the analysis of the samples from same lot under a variety of normal test
conditions (i.e. different laboratories, analysts, days & Instruments).
The ruggedness of assay method of Guaiphensin in Bronsyp Syrup shall be studied by
analyzing the samples from same lot by two different analysts from same laboratory or at two
different laboratories by two different analysts on two different days.
RUGGEDNESS STUDY DATA
Ruggedness data for assay method by two different analysts on different days and different instrument.
Sr. No.
Analyst- I (Satpal Saini)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Analyst- II (Rajnish Kohli)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Wt. of Ref.
Standard gm.
Wt. of Sample
ml
Measured
Absor-bance
at of Ref.
Std.
Measured Absor-
bance at 275nm of
sample
% of Guaiphenesin
Wt. of Ref. Standard
gm.
Wt. of Sample
ml
Measured Absor-
bance at of Ref.
Std.
Measured Absor-
bance at 275nm
of sample
% of Guaiphen
esin
01
02
03
Label Claim 50mg Guaiphenesin/5ml Label Claim 50mg Guaiphenesin/5ml
Mean Mean
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : Standard Deviation : ±10 %
Sarthak Biotech Pvt. Ltd. Total Page: 01-16Assay Method Validation QC/VLD/11Name of the Product: BECLOSAR-NC CREAMPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 08/02/2008 Review Date : 07/02/2010 Revision No. : 00
PURPOSE : The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Beclomethasone Dipropionate, Neomycin Sulphate & Clotrimazole in Beclosar-NC Cream will give results with acceptable Accuracy, Linearity, Repeatibility, Relative Standard Deviation & Ruggedness when applied to the Beclosar-NC Cream.
SCOPE : This method is applies to the procedure which is used for calculation and assay of Beclomethasone Dipropionate, Neomycin Sulphate & Clotrimazole.
RESPONSIBILITY: 1. Quality Control Manager.
2. Asstt. Quality Control Chemist.
VALIDATION PARAMETERS:
1. Linearity.
2. Repeatibility.
3. Relative Standard Deviation.
4. Ruggedness.
Sarthak Biotech Pvt. Ltd. Total Page: 02-16Assay Method Validation QC/VLD/11Name of the Product: BECLOSAR-NC CREAMPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 08/02/2008 Review Date : 07/02/2010 Revision No. : 00
METHOD : (Beclomethasone Dipropionate)
1. Weigh accurately 50mg. of Beclomethasone Dipropionate and dissolve in 100ml. of methanol. Further dilute 1ml. and make up the volume to 50ml. with methanol. This makes 10mcg./ml dilution of Beclomethasone Dipropionate.
2. Weight accurately sample (cream) equivalent to 0.5mg of Beclomethasone Dipropionate and dissolve in 25ml. of methanol. Filter and then make up the volume to 50ml. with methanol.
3. Pipette 1ml, 2ml, 3ml, 4ml & 5ml of standard and sample solution, add 6ml. of 0.1M
tetrabutyl ammonium hydroxide (TBAH) reagent and 2ml. of 0.5%w/v of Tetrazolium blue (BTZ) solution. Heat the solution on water bath for 15minutes, cool the solution and add 10ml. Glacial Acetic acid.
4. Measure the absorbance without delay at maximum at about 515nm, against respective
blank and tabulate the observed data in the observation table.
5. Plot linearity curve from the different concentration corresponding area of Beclomethasone Dipropionate peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.
6. Plot repeatability curve from by taking Beclomethasone Dipropionate solution serial no. on X-axis and observed peak area on Y-axis.
7. Draw a line which should cover almost all the points. The resulted graph should be
almost parallel to the X-axis. . .
8. Calculated the result by comparison with peak area obtained in standard and sample solution respectively.
Sarthak Biotech Pvt. Ltd. Total Page: 03-16Assay Method Validation QC/VLD/11Name of the Product: BECLOSAR-NC CREAMPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 08/02/2008 Review Date : 07/02/2010 Revision No. : 00
LINEARITY STUDY
The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range.
Linearity of assay method of Beclomethasone Dipropionate in Beclosar-NC Cream shall be studied by measuring the absorbance of the standard Beclomethasone Dipropionate in solution of 1 – 5mcg/ml concentration & range 515nm. Linearity curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR LINEARITY
Sr.No. Concentration of Beclomethasone Dipropionate in mcg\ml Peak area of
1. 1mcg\ml 680
2. 2mcg\ml 742
3. 3mcg\ml 791
4. 4mcg\ml 839
5. 5mcg\ml 872
Sarthak Biotech Pvt. Ltd. Total Page: 04-16Assay Method Validation QC/VLD/11Name of the Product: BECLOSAR-NC CREAMPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 08/02/2008 Review Date : 07/02/2010 Revision No. : 00
REPEATIBILITY STUDY
Repeatibility of assay method of Beclomethasone Dipropionate in Beclosar-NC Cream shall be studied by measuring the absorbance of the standard Beclomethasone Dipropionate in solution of 1 mcg/ml concentration & range 515nm. Repeatibility curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR REPEATIBILITY
Sr.No. Concentration in mcg\ml Peak area
1 1mcg\ml 680
2 . 1mcg\ml 682
3. 1mcg\ml 681
4. 1mcg\ml 680
5. 1mcg\ml 685
Sarthak Biotech Pvt. Ltd. Total Page: 05-16Assay Method Validation QC/VLD/11Name of the Product: BECLOSAR-NC CREAMPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 08/02/2008 Review Date : 07/02/2010 Revision No. : 00
RELATIVE STANDARD DEVIATION Test concentration of Beclomethasone Dipropionate used = 1mcg\ml. Number of test solution used = 5 680+682+681+680+685 Average area = ----------------------------------- = 681.6
5
(680-681.6)2 +(682-681.6)2 +(681-681.6)2 +(680-681.6)2 +(685-681.6)2
Dev. S = --------------------------------------------------------------------------------------- 1/2
5-1
2.56+0.16+0.36+2.56+11.56 = ---------------------------------------1/2
4 17.2 = ----- ----1/2
4 = 4.3 1/2
= 2.0736
2.0736 x 100 RSD = ------------------ = 0.3042 %
681.6
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd. Total Page: 06-16Assay Method Validation QC/VLD/11Name of the Product: BECLOSAR-NC CREAMPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 08/02/2008 Review Date : 07/02/2010 Revision No. : 00
RUGGEDNESS STUDY
The ruggedness of an analytical method is the degree of reproducibility of the test
results obtained by the analysis of the samples from same lot under a variety of normal test
conditions (i.e. different laboratories, analysts, days & Instruments).
The ruggedness of assay method of Beclomethasone Dipropionate in Beclosar-NC
Cream shall be studied by analyzing the samples from same lot by two different analysts from
same laboratory or at two different laboratories by two different analysts on two different days.
RUGGEDNESS STUDY DATA
Ruggedness data for assay method by two different analysts on different days and different instrument.
Sr. No.
Analyst- I (Satpal Saini)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Analyst- II (Rajnish Kohli)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Wt. of Ref.
Standardgm.
Wt. of Sample
gm
Measured
Absor-bance
at of Ref.
Std.
Measured Absor-
bance at 515nm of
sample
% of Beclomethasone
Wt. of Ref.
Standardgm.
Wt. of Sample
ml
Measured Absor-
bance at of Ref.
Std.
Measured Absor-
bance at 515nm
of sample
% of Beclomet
hasone
01 0.0498 1.9969 683 681 99.52 0.0506 1.9978 682 684 100.62
02 0.0492 2.0013 685 686 98.53 0.0494 1.9982 682 684 99.19
03 0.0502 1.9975 684 686 100.88 0.0497 2.0018 684 686 99.61
Label Claim 0.025% w/w Label Claim 0.025% w/w
Mean 99.64% Mean 100.14%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 99.89 %Standard Deviation : ±10%
Sarthak Biotech Pvt. Ltd. Total Page: 07-16Assay Method Validation QC/VLD/11Name of the Product: BECLOSAR-NC CREAMPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 08/02/2008 Review Date : 07/02/2010 Revision No. : 00
METHOD : (Neomycin Sulphate)
1. Weigh accurately 100mg. of Neomycin Sulphate and dissolve in 100ml. of phosphate buffer pH 8.0 and prepare the dilution 100mcg/ml. in phosphate buffer.
2. Weight accurately sample (cream) equivalent to 10mg of Neomycin Sulphate, suspend
in 25ml. of phosphate buffer and extract with Chloroform (3 x 30ml). Collect chloroform in a dry 100ml volumetric flask filtering each layer through same anhydrous sodium Sulphate bed. Rinse sodium Sulphate layer with chloroform, the combined chloroform layer is made up of volume. Aqueous buffered layer heated in water bath till no perceptible smell of chloroform is diluted to 100ml with phosphate buffer and used for estimation of Neomycin Sulphate.
3. Pipette 5ml, 6ml, 7ml, 8ml & 9ml of standard and sample solution, add 5ml. of freshly prepared ninhydrin reagent and heat in water bath for 20 minutes, cool, add 2 ml. of methanol and dilute to 50ml with phosphate buffer.
4. Measure the absorbance without delay at maximum at about 570nm, against respective
blank and tabulate the observed data in the observation table.
5. Plot linearity curve from the different concentration corresponding area of Neomycin Sulphate peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.
6. Plot repeatability curve from by taking Neomycin Sulphate solution serial no. on X-axis and observed peak area on Y-axis.
7. Draw a line which should cover almost all the points. The resulted graph should be
almost parallel to the X-axis. . .
8. Calculated the result by comparison with peak area obtained in standard and sample solution respectively.
Sarthak Biotech Pvt. Ltd. Total Page: 08-16Assay Method Validation QC/VLD/11Name of the Product: BECLOSAR-NC CREAMPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 08/02/2008 Review Date : 07/02/2010 Revision No. : 00
LINEARITY STUDY
The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range.
Linearity of assay method of Neomycin Sulphate in Beclosar-NC Cream shall be studied by measuring the absorbance of the standard Neomycin Sulphate in solution of 10 – 18mcg/ml concentration & range 570nm. Linearity curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR LINEARITY
Sr.No. Concentration of Neomycin Sulphate in mcg\ml Peak area of
1. 10mcg\ml 652
2. 12mcg\ml 698
3. 14mcg\ml 749
4. 16mcg\ml 805
5. 18mcg\ml 862
Sarthak Biotech Pvt. Ltd. Total Page: 09-16Assay Method Validation QC/VLD/11Name of the Product: BECLOSAR-NC CREAMPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 08/02/2008 Review Date : 07/02/2010 Revision No. : 00
REPEATIBILITY STUDY
Repeatibility of assay method of Neomycin Sulphate in Beclosar-NC Cream shall be studied by measuring the absorbance of the standard Neomycin Sulphate in solution of 10mcg/ml concentration & range 570nm. Repeatibility curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR REPEATIBILITY
Sr.No. Concentration in mcg\ml Peak area
1 10mcg\ml 650
2 . 10mcg\ml 652
3. 10mcg\ml 654
4. 10mcg\ml 648
5. 10mcg\ml 657
Sarthak Biotech Pvt. Ltd. Total Page: 10-16Assay Method Validation QC/VLD/11Name of the Product: BECLOSAR-NC CREAMPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 08/02/2008 Review Date : 07/02/2010 Revision No. : 00
RELATIVE STANDARD DEVIATION
Test concentration of Neomycin Sulphate used = 10mcg\ml. Number of test solution used = 5 650+652+654+648+657 Average area = ----------------------------------- = 652.2
5
(650-652.2)2 +(652-652.2)2 +(654-652.2)2 +(648-652.2)2 +(657-652.2)2
Dev. S = --------------------------------------------------------------------------------------- 1/2
5-1
4.84+0.04+3.24+17.64+23.04 = ---------------------------------------1/2
4 48.8 = ----- ----1/2
4 = 12.2 1/2
= 3.493
3.493 x 100 RSD = ------------------ = 0.536 %
652.2
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd. Total Page: 11-16Assay Method Validation QC/VLD/11Name of the Product: BECLOSAR-NC CREAMPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 08/02/2008 Review Date : 07/02/2010 Revision No. : 00
RUGGEDNESS STUDY
The ruggedness of an analytical method is the degree of reproducibility of the test
results obtained by the analysis of the samples from same lot under a variety of normal test
conditions (i.e. different laboratories, analysts, days & Instruments).
The ruggedness of assay method of Neomycin Sulphate in Beclosar-NC Cream shall
be studied by analyzing the samples from same lot by two different analysts from same
laboratory or at two different laboratories by two different analysts on two different days.
RUGGEDNESS STUDY DATA
Ruggedness data for assay method by two different analysts on different days and different instrument.
Sr. No.
Analyst- I (Satpal Saini)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Analyst- II (Rajnish Kohli)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Wt. of Ref.
Standardgm.
Wt. of Sample
gm.
Measured
Absor-bance
at of Ref.
Std.
Measured Absor-
bance at 570nm of
sample
% of Neomyci
n
Wt. of Ref.
Standardgm.
Wt. of Sample
gm.
Measured Absor-
bance at of Ref.
Std.
Measured Absor-
bance at 570nm
of sample
% of Neomyci
n
01 0.0998 2.0256 656 652 98.03 0.0995 2.0160 652 649 98.37
02 0.0995 2.0106 650 652 99.37 0.0998 2.0059 653 654 99.71
03 0.1008 1.9994 651 649 100.62 0.1029 1.9989 656 658 103.33
Label Claim 0.5% w/w Label Claim 0.5% w/w
Mean 99.34% Mean 100.45 %
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 99.89%Standard Deviation : ±10
Sarthak Biotech Pvt. Ltd. Total Page: 12-16Assay Method Validation QC/VLD/11Name of the Product: BECLOSAR-NC CREAMPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 08/02/2008 Review Date : 07/02/2010 Revision No. : 00
METHOD : (Clotrimazole)
1. Weigh accurately reference standard 100mg. of Clotrimazole and dissolve in 100ml. of 0.1M hydrochloric acid and prepare the dilution 100mcg./ml of Clotrimazole.
2. Weigh accurately sample equivalent to 100mg. of Clotrimazole and dissolve in 100ml.
of 0.1M hydrochloric acid and prepare the dilution 100mcg./ml of Clotrimazole.
3. Pipette 1ml, 2ml, 3ml, 4ml & 5ml of standard and sample dilution, add 3ml. of citric acid solution and 2ml of dye solution. Extract the colour with three 8 ml portions of chloroform. Make up the volume to 25ml and pass through anhydrous sodium Sulphate.
4. Measure the absorbance without delay at maximum at about 422nm, against respective
blank and tabulate the observed data in the observation table.
5. Plot linearity curve from the different concentration corresponding area of Clotrimazole peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.
6. Plot repeatability curve from by taking Clotrimazole solution serial no. on X-axis and observed peak area on Y-axis.
7. Draw a line which should cover almost all the points. The resulted graph should be
almost parallel to the X-axis. . .
8. Calculated the result by comparison with peak area obtained in standard and sample solution respectively.
Sarthak Biotech Pvt. Ltd. Total Page: 13-16Assay Method Validation QC/VLD/11Name of the Product: BECLOSAR-NC CREAMPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 08/02/2008 Review Date : 07/02/2010 Revision No. : 00
LINEARITY STUDY
The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range.
Linearity of assay method of Clotrimazole in Beclosar-NC Cream shall be studied by measuring the absorbance of the standard Clotrimazole in solution of 4 – 20mcg/ml concentration & range 422nm. Linearity curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR LINEARITY
Sr.No. Concentration of Clotrimazole in mcg\ml Peak area of 1. 4mcg\ml 595
2. 8mcg\ml 632
3. 12mcg\ml 682
4. 16mcg\ml 735
5. 20mcg\ml 795
Sarthak Biotech Pvt. Ltd. Total Page: 14-16Assay Method Validation QC/VLD/11Name of the Product: BECLOSAR-NC CREAMPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 08/02/2008 Review Date : 07/02/2010 Revision No. : 00
REPEATIBILITY STUDY
Repeatibility of assay method of Clotrimazole in Beclosar-NC Cream shall be studied by measuring the absorbance of the standard Clotrimazole in solution of 4mcg/ml concentration & range 422nm. Repeatibility curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR REPEATIBILITY
Sr.No. Concentration in mcg\ml Peak area
1 4mcg\ml 595
2 . 4mcg\ml 592
3. 4mcg\ml 593
4. 4mcg\ml 595
5. 4mcg\ml 596
Sarthak Biotech Pvt. Ltd. Total Page: 15-16Assay Method Validation QC/VLD/11Name of the Product: BECLOSAR-NC CREAMPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 08/02/2008 Review Date : 07/02/2010 Revision No. : 00
RELATIVE STANDARD DEVIATION
Test concentration of Neomycin Sulphate used = 4mcg\ml. Number of test solution used = 5 595+592+593+595+596 Average area = ----------------------------------- = 594.2
5
(595-594.2)2 +(592-594.2)2 +(593-594.2)2 +(595-594.2)2 +(596-594.2)2
Dev. S = --------------------------------------------------------------------------------------- 1/2
5-1
0.64 + 4.84 + 1.44 + 0.64 + 3.24 = ------------------------------------------1/2
4 10.8 = ----- ----1/2
4 = 2.7 1/2
= 1.6431
1.6431 x 100 RSD = ------------------ = 0.276 %
594.2
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd. Total Page: 16-16Assay Method Validation QC/VLD/11Name of the Product: BECLOSAR-NC CREAMPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 08/02/2008 Review Date : 07/02/2010 Revision No. : 00
RUGGEDNESS STUDY
The ruggedness of an analytical method is the degree of reproducibility of the test
results obtained by the analysis of the samples from same lot under a variety of normal test
conditions (i.e. different laboratories, analysts, days & Instruments).
The ruggedness of assay method of Clotrimazole in Beclosar-NC Cream shall be
studied by analyzing the samples from same lot by two different analysts from same laboratory
or at two different laboratories by two different analysts on two different days.
RUGGEDNESS STUDY DATA
Ruggedness data for assay method by two different analysts on different days and different instrument.
Sr. No.
Analyst- I (Rajnish Kohli)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Analyst- II (Satpal Saini)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Wt. of Ref.
Standardgm
Wt. of Sample gm
Measured Absor-
bance at of Ref.
Std.
Measured
Absor-bance at 422nm
of sample
% of Clotrima
zole
Wt. of Ref.
Standardgm
Wt. of Sample
gm
Measured Absor-
bance at of Ref.
Std.
Measured Absor-
bance at 422nm
of sample
% of Clotrimaz
ole
01 0.0997 10.0056 596 598 100.03 0.1002 9.9969 593 595 100.57
02 0.1005 9.9969 592 593 100.76 0.0998 9.9982 598 595 99.32
03 0.9989 10.0125 593 590 99.32 0.1012 10.0015 597 599 101.53
Label Claim 1% w/w Label Claim 1% w/w
Mean 100.03% Mean 100.47%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 100.25%Standard Deviation : ±10%
Sarthak Biotech Pvt. Ltd. Total Page: 01-11Assay Method Validation QC/VLD/12Name of the Product: MOVSAR GELPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 04/11/2007 Review Date : 03/11/2009 Revision No. : 00
PURPOSE : The establish documented evidence which will provide a high degree of assurance that the analytical method for assay of Diclofenac Diethylamine & Methylsalicylate in Movsar Gel will give results with acceptable Accuracy, Linearity, Repeatibility, Relative Standard Deviation & Ruggedness when applied to the Movsar Gel.
SCOPE : This method is applies to the procedure which is used for calculation and assay of Diclofenac diethylamine & Methylsalicylate.
RESPONSIBILITY: 1. Quality Control Manager.
2. Asstt. Quality Control Chemist.
VALIDATION PARAMETERS:
1. Linearity.
2. Repeatibility.
3. Relative Standard Deviation.
4. Ruggedness.
Sarthak Biotech Pvt. Ltd. Total Page: 02-11Assay Method Validation QC/VLD/12Name of the Product: MOVSAR GELPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 04/11/2007 Review Date : 03/11/2009 Revision No. : 00
METHOD : (Diclofenac diethylamine )
1. Weigh accurately 50mg. of Diclofenac sodium working standard in a 100ml volumetric flask. Dissolve with phosphate buffer and make up to 100ml with same solution and prepared the dilution 50mcg./ml. of Diclofenac sodium.
2. Weight accurately sample equivalent to 5mg of Diclofenac sodium in a 100ml volumetric flask and dissolve in 100ml phosphate buffer.
3. Pipette 10ml, 12ml, 15ml, 18ml & 20ml of standard and sample solution separately to
each funnel add 5ml of methylene blue solution and extract with (10+10+5ml) of chloroform passing each extract through anhydrous sodium Sulphate, dilute to 25ml with chloroform mix to make homogeneous.
4. Measure the absorbance without delay at maximum at about 650nm, against respective blank and tabulate the observed data in the observation table.
5. Plot linearity curve from the different concentration corresponding area of Diclofenac diethylamine peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.
6. Plot repeatability curve from by taking Diclofenac diethylamine solution serial no. on X-axis and observed peak area on Y-axis.
7. Draw a line which should cover almost all the points. The resulted graph should be
almost parallel to the X-axis. . .
8. Calculated the result by comparison with peak area obtained in standard and sample solution respectively.
Sarthak Biotech Pvt. Ltd. Total Page: 03-11Assay Method Validation QC/VLD/12Name of the Product: MOVSAR GELPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 04/11/2007 Review Date : 03/11/2009 Revision No. : 00
LINEARITY STUDY
The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range.
Linearity of assay method of Diclofenac diethylamine in Movsar Gel shall be studied by measuring the absorbance of the standard Diclofenac diethylamine in solution of 20 – 40mcg/ml concentration & range 650nm. Linearity curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR LINEARITY
Sr.No. Concentration of Diclofenac diethylamine in mcg\ml Peak area of 1. 20mcg\ml 716
2. 24mcg\ml 742
3. 30mcg\ml 795
4. 36mcg\ml 842
5. 40mcg\ml 878
Sarthak Biotech Pvt. Ltd. Total Page: 04-11Assay Method Validation QC/VLD/12Name of the Product: MOVSAR GELPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 04/11/2007 Review Date : 03/11/2009 Revision No. : 00
REPEATIBILITY STUDY
Repeatibility of assay method of Diclofenac diethylamine in Movsar Gel shall be studied by measuring the absorbance of the standard Diclofenac diethylamine in solution of 20 mcg/ml concentration & range 650nm. Repeatibility curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR REPEATIBILITY
Sr.No. Concentration in mcg\ml Peak area
1 20mcg\ml 719
2 . 20mcg\ml 714
3. 20mcg\ml 718
4. 20mcg\ml 722
5. 20mcg\ml 720
Sarthak Biotech Pvt. Ltd. Total Page: 05-11Assay Method Validation QC/VLD/12Name of the Product: MOVSAR GELPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 04/11/2007 Review Date : 03/11/2009 Revision No. : 00
RELATIVE STANDARD DEVIATION
Test concentration of Diclofenac diethylamine used = 20mcg\ml. Number of test solution used = 5 719+714+718+722+720 Average area = ----------------------------------- = 718.6
5
(719-718.6)2 +(714-718.6)2 +(718-718.6)2 +(722-718.6)2 +(720-718.6)2
Dev. S = --------------------------------------------------------------------------------------- 1/2
5-1
0.16+21.16+ 0.36+ 11.56+ 1.96 = ------------------------------------------1/2
4
35.2 = ----- ----1/2
4 = 8.8 1/2
= 2.9664
2.9664 x 100 RSD = ------------------ = 0.412 %
718.6
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd. Total Page: 06-11Assay Method Validation QC/VLD/12Name of the Product: MOVSAR GELPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 04/11/2007 Review Date : 03/11/2009 Revision No. : 00
RUGGEDNESS STUDY
The ruggedness of an analytical method is the degree of reproducibility of the test
results obtained by the analysis of the samples from same lot under a variety of normal test
conditions (i.e. different laboratories, analysts, days & Instruments).
The ruggedness of assay method of Diclofenac diethylamine in Movsar Gel shall be
studied by analyzing the samples from same lot by two different analysts from same laboratory
or at two different laboratories by two different analysts on two different days.
RUGGEDNESS STUDY DATA
Ruggedness data for assay method by two different analysts on different days and different instrument.
Sr. No.
Analyst- I (Rajnish Kohli)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Analyst- II (Satpal Saini)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Wt. of Ref.
Standardgm
Wt. of Sample
gm.
Measured Absorba
nce at of Ref.
Std.
Measured Absorbanc
e at 650nm of
sample
% of Diclofenac
diethylamine
Wt. of Ref.
Standardgm.
Wt. of Sample
gm.
Measured Absor-
bance at of Ref.
Std.
Measured Absor-
bance at 650nm
of sample
% of Diclofe
nac diethylamine
01 0.0493 0.5012 715 718 99.78 0.0498 0.4992 718 720 100.03
02 0.0501 0.5006 718 716 99.80 0.0494 0.4984 716 718 99.39
03 0.0499 0.4993 719 716 99.52 0.0502 0.5012 719 721 100.44
Label Claim 1.16% w/w Label Claim 1.16% w/w
Mean 99.36% Mean 100.44
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 99.88 %Standard Deviation : ±10%
Sarthak Biotech Pvt. Ltd. Total Page: 07-11Assay Method Validation QC/VLD/12Name of the Product: MOVSAR GELPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 04/11/2007 Review Date : 03/11/2009 Revision No. : 00
METHOD : (Methylsalicylate)
1. Weigh accurately 100mg. of Methylsalicylate as working standard in a 100ml volumetric flask and dissolve in 0.02M hydrochloric acid and make up to produced 100ml, again dilute 15ml of the resulting solution to 100ml with 0.02M hydrochloric acid.
2. Weight accurately equivalent to 100mg of Methyl salicylate in a 500ml conical flask
add 30ml of 0.5M sodium hydroxide and 20ml of methyl alcohol, mix and reflux for one hour on steam bath to sponify and hydrolyse, cool to room temperature and add 20ml of 1.0M hydrochloric acid and dilute to 100ml with water and filter. Further dilute 15ml to 100ml with 0.02M hydrochloric acid.
3. Pipette 10ml, 15ml, 20ml, 25ml & 30ml of standard and sample solution in separately
funnel and extract with 25ml of chloroform, separate chloroform layer in a dry test tube. To 5 ml of chloroform layer add 10ml of ferric ammonium sulfate reagent and separate the aqueous layer.
4. Measure the absorbance without delay at maximum at about 530nm, against respective
blank and tabulate the observed data in the observation table.
5. Plot linearity curve from the different concentration corresponding area of Methylsalicylate peak. Taking concentration on X-axis and corresponding peak on Y-axis. Plot graph should be connected at least three points.
6. Plot repeatability curve from by taking Methylsalicylate solution serial no. on X-axis and observed peak area on Y-axis.
7. Draw a line which should cover almost all the points. The resulted graph should be
almost parallel to the X-axis. . .
8. Calculated the result by comparison with peak area obtained in standard and sample solution respectively.
Sarthak Biotech Pvt. Ltd. Total Page: 08-11Assay Method Validation QC/VLD/12Name of the Product: MOVSAR GELPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 04/11/2007 Review Date : 03/11/2009 Revision No. : 00
LINEARITY STUDY
The linearity of an analytical method is its ability to elicit test results data directly proportional to the concentration of the analyte in samples within given range.
Linearity of assay method of Methylsalicylate in Movsar Gel shall be studied by measuring the absorbance of the standard Methylsalicylate in solution of 30 – 90mcg/ml concentration & range 530nm. Linearity curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR LINEARITY
Sr.No. Concentration of Methylsalicylate in mcg\ml Peak area 1. 30mcg\ml 685
2. 45mcg\ml 752
3. 60mcg\ml 796
4. 75mcg\ml 855
5. 90mcg\ml 888
Sarthak Biotech Pvt. Ltd. Total Page: 09-11Assay Method Validation QC/VLD/12Name of the Product: MOVSAR GELPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 04/11/2007 Review Date : 03/11/2009 Revision No. : 00
REPEATIBILITY STUDY
Repeatibility of assay method of Methylsalicylate in Movsar Gel shall be studied by measuring the absorbance of the standard Methylsalicylate in solution of 30mcg/ml concentration & range 530nm. Repeatibility curve shall be plotted for absorbance against concentration.
OBSERVATION TABLE FOR REPEATIBILITY
Sr.No. Concentration in mcg\ml Peak area
1 30mcg\ml 687
2 . 30mcg\ml 685
3. 30mcg\ml 684
4. 30mcg\ml 686
5. 30mcg\ml 685
Sarthak Biotech Pvt. Ltd. Total Page: 10-11Assay Method Validation QC/VLD/12Name of the Product: MOVSAR GELPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 04/11/2007 Review Date : 03/11/2009 Revision No. : 00
RELATIVE STANDARD DEVIATION
Test concentration of Methylsalicylate used = 30mcg\ml. Number of test solution used = 5 687+685+684+686+685 Average area = ----------------------------------- = 685.4
5
(687-685.4)2 +(685-685.4)2 +(684-685.4)2 +(686-685.4)2 +(685-685.4)2
Dev. S = --------------------------------------------------------------------------------------- 1/2
5-1
2.56+0.16+1.96+0.36+0.16 = ----------------------------------------1/2
4 = 5.2
[ ----- ----1/2
4
= 1.3 1/2
= 1.1401
1.1401 x 100 RSD = ------------------ = 0.166 %
685.4
Limit : ( Not more than 2.0% )
Sarthak Biotech Pvt. Ltd. Total Page: 11-11Assay Method Validation QC/VLD/12Name of the Product: MOVSAR GELPrepared By Checked By/ Approved By
Date of Issue/Effective Date : 04/11/2007 Review Date : 03/11/2009 Revision No. : 00
RUGGEDNESS STUDY
The ruggedness of an analytical method is the degree of reproducibility of the test
results obtained by the analysis of the samples from same lot under a variety of normal test
conditions (i.e. different laboratories, analysts, days & Instruments).
The ruggedness of assay method of Methylsalicylate in Movsar Gel shall be studied
by analyzing the samples from same lot by two different analysts from same laboratory or at
two different laboratories by two different analysts on two different days.
RUGGEDNESS STUDY DATA
Ruggedness data for assay method by two different analysts on different days and different instrument.
Sr. No.
Analyst- I (Satpal Saini)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Analyst- II (Rajnish Kohli)Date of analysis :Instrument & Model : UV Spectrophotometer, Systronics 117Location : Instrument Lab.
Wt. of Ref.
Standardgm
Wt. of Sample
gm
Measured
Absorbance at
of Ref. Std.
Measured Absorbance
at 530nm of sample
% of Methylsalicylate
Wt. of Ref.
Standardgm
Wt. of Sample
gm.
Measured Absorbanc
e at of Ref.
Std.
Measured Absorban
ce at 530nm
of sample
% of Methylsal
icylate
01 0.0998 0.9994 684 686 100.15 0.1018 0.9996 687 686 101.68
02 0.1009 0.9989 684 685 100.87 0.0998 1.0015 684 686 99.93
03 0.0998 1.0012 687 685 99.39 0.1009 0.9994 684 682 100.66
Label Claim 10% w/w Label Claim 10% w/w
Mean 100.31 % Mean 100.75%
Standard Deviation 10% Standard Deviation 10%
Combining mean of I & II of both the analysts : Mean : 100.53% Standard Deviation : ±10%